Sauerkraut study - Regression-based Screening on functional data (on genes, not pathways)

Code

library(sauerkrautTaxonomyBuddy)
library(SummarizedExperiment) # microbiome analysis
library(mia)                  # microbiome analysis

library(readxl)     # read Excel files
library(mgcv)       # regression model
library(NBZIMM)     # zero-inflated Gaussian mixed regression models
library(parallel)   # parallelize processes on multiple cores

library(dplyr)      # data handling
library(tidyr)      # data transformation
library(ggplot2)    # data visualization
library(patchwork)  # grid of ggplot2
library(ggpubr)     # further ggplot2 grid customizations
library(kableExtra) # print tables

# set ggplot2 theme
theme_set(
  theme_minimal() +
    theme(plot.title       = element_text(hjust = 0.5),
          plot.subtitle    = element_text(hjust = 0.5),
          panel.grid.minor = element_blank(),
          plot.background  = element_rect(fill = "white", color = "white"))
)

# suppress scientific notation (= e notation of decimal numbers)
options(scipen = 999)

Code

# read the Excel file from Till Robin, mapping individual genes to specific SCFAs
path_SCFAgenes <- "9_lookup_SCFA-KEGG-KO.xlsx"
dat_scfaLookup <- read_excel(path_SCFAgenes, sheet = 1) %>% dplyr::select(SCFA, KO)

Data preparation

Code

define_dataPaths()

Code

# KO data
tse_KO <- readAndPrepare_mainFunctionalKOdata(
  path_KEGG_KO                      = path_KEGGKO_funData,
  path_KEGG_pathways                = path_KEGGPathways_funData,
  path_groupInfoRdata               = path_groupInfoRdata,
  path_participantRdata             = path_participantRdata,
  path_samplesIDxlsx                = path_samplesIDxlsx,
  path_dietInfoCsv                  = path_dietInfoCsv,
  path_nutrientInfoCsv              = path_nutrientInfoCsv,
  path_bloodMetabolomeXlsx          = path_bloodMetabolomeXlsx,
  path_stoolMetabolomeXlsx          = path_stoolMetabolomeXlsx,
  path_T4questXlsx                  = path_T4questXlsx,
  path_stoolInfoXlsx                = path_stoolInfoXlsx,
  path_bodyMeasuresXlsx             = path_bodyMeasuresXlsx,
  path_sampleLookupXlsx             = path_sampleLookupXlsx,
  path_krautLookupXlsx              = path_krautLookupXlsx,
  path_bloodMarkerBostonRdata       = path_bloodMarkerBostonRdata,
  path_bloodMarkerZentrallaborRdata = path_bloodMarkerZentrallaborRdata,
  path_bloodMarkerBostonXlsx        = path_bloodMarkerBostonXlsx,
  path_bloodMarkerZentrallaborXlsx  = path_bloodMarkerZentrallaborXlsx,
  exclude_sickParticipants          = "taxonomy paper"
)

# pathway data
tse_PW <- readAndPrepare_mainFunctionalPathwayData(
  path_KEGG_KO                      = path_KEGGKO_funData,
  path_KEGG_pathways                = path_KEGGPathways_funData,
  path_groupInfoRdata               = path_groupInfoRdata,
  path_participantRdata             = path_participantRdata,
  path_samplesIDxlsx                = path_samplesIDxlsx,
  path_dietInfoCsv                  = path_dietInfoCsv,
  path_nutrientInfoCsv              = path_nutrientInfoCsv,
  path_bloodMetabolomeXlsx          = path_bloodMetabolomeXlsx,
  path_stoolMetabolomeXlsx          = path_stoolMetabolomeXlsx,
  path_T4questXlsx                  = path_T4questXlsx,
  path_stoolInfoXlsx                = path_stoolInfoXlsx,
  path_bodyMeasuresXlsx             = path_bodyMeasuresXlsx,
  path_sampleLookupXlsx             = path_sampleLookupXlsx,
  path_krautLookupXlsx              = path_krautLookupXlsx,
  path_bloodMarkerBostonRdata       = path_bloodMarkerBostonRdata,
  path_bloodMarkerZentrallaborRdata = path_bloodMarkerZentrallaborRdata,
  path_bloodMarkerBostonXlsx        = path_bloodMarkerBostonXlsx,
  path_bloodMarkerZentrallaborXlsx  = path_bloodMarkerZentrallaborXlsx,
  exclude_sickParticipants          = "taxonomy paper"
)

# create joint tables with all relevant information
dat_KO <- tse_KO %>% mia::meltAssay(assay.type = "tpmAbd", add_row_data = TRUE, add_col_data = TRUE)
dat_PW <- tse_PW %>% mia::meltAssay(assay.type = "tpmAbd", add_row_data = TRUE, add_col_data = TRUE)

Code

# save the average T1/T3 baseline Shannon diversity as individual variable
tse <- readAndPrepare_mainTaxonomicData(path_groupInfoRdata               = path_groupInfoRdata,
                                        path_participantRdata             = path_participantRdata,
                                        path_samplesIDxlsx                = path_samplesIDxlsx,
                                        path_dietInfoCsv                  = path_dietInfoCsv,
                                        path_nutrientInfoCsv              = path_nutrientInfoCsv,
                                        path_bloodMetabolomeXlsx          = path_bloodMetabolomeXlsx,
                                        path_stoolMetabolomeXlsx          = path_stoolMetabolomeXlsx,
                                        path_T4questXlsx                  = path_T4questXlsx,
                                        path_stoolInfoXlsx                = path_stoolInfoXlsx,
                                        path_bodyMeasuresXlsx             = path_bodyMeasuresXlsx,
                                        path_absTaxonomyData              = path_absTaxonomyData,
                                        path_relTaxonomyData              = path_relTaxonomyData,
                                        path_sampleLookupXlsx             = path_sampleLookupXlsx,
                                        path_krautLookupXlsx              = path_krautLookupXlsx,
                                        path_bloodMarkerBostonRdata       = path_bloodMarkerBostonRdata,
                                        path_bloodMarkerZentrallaborRdata = path_bloodMarkerZentrallaborRdata,
                                        path_bloodMarkerBostonXlsx        = path_bloodMarkerBostonXlsx,
                                        path_bloodMarkerZentrallaborXlsx  = path_bloodMarkerZentrallaborXlsx,
                                        aggregation_level                 = "species",
                                        exclude_sickParticipants          = "taxonomy paper")

Only retaining stool bacteria with a minimum abundance of 0.01% in at least 10% of samples...

Warning in toTree(td): The root is added with label 'ALL'

Warning: The 'getPrevalentTaxa' function is deprecated. Use
'getPrevalentFeatures' instead.

Code

dat <- tse %>% 
  colData() %>% 
  as.data.frame()

dat_baselineDiv <- dat %>% 
  filter(timepoint %in% c("T1", "T3")) %>% 
  group_by(participant_id) %>% 
  summarize(baseline_div = mean(diversity_shannon, na.rm = TRUE))
dat_KO <- dat_KO %>% dplyr::left_join(dat_baselineDiv, by = "participant_id")
dat_PW <- dat_PW %>% dplyr::left_join(dat_baselineDiv, by = "participant_id")

Code

# prepare data for regression models
median_baselineDiv <- median(dat_baselineDiv$baseline_div)
dat_KO <- dat_KO %>% 
  filter(intervention != "FollowUp") %>% 
  mutate(tpmAbd_log     = log(tpmAbd + 1),
         intervention   = case_when(treatment == "Baseline" ~ "Baseline",
                                    TRUE                    ~ intervention),
         age_centered50 = age - 50,
         bmi_centered25 = bmi_t0 - 25,
         age_cat        = case_when(age < 50 ~ paste0(min(age), "-49"),
                                    TRUE     ~ paste0("50-", max(age))),
         age_cat        = factor(age_cat),
         bmi_cat        = case_when(bmi_t0 < 25 ~ paste0("[", floor(min(bmi_t0)), ", 25)"),
                                    TRUE        ~ paste0("[25, ", ceiling(max(bmi_t0)), "]")),
         bmi_cat        = factor(bmi_cat),
         phase          = case_when(timepoint %in% c("T3","T4") ~ "phase 2",
                                    TRUE                        ~ "phase 1"),
         phase          = factor(phase),
         dailyFiber_cat  = case_when(dailyConsumption_fiber >= 30 ~ ">= 30g",
                                     TRUE                         ~ "< 30g"),
         dailyFiber_cat  = factor(dailyFiber_cat),
         baselineDiv_cat = case_when(baseline_div < median_baselineDiv ~ "[2.8, 3.85)",
                                     TRUE                              ~ "[3.85, 4.4)"),
         baselineDiv_cat = factor(baselineDiv_cat))

dat_PW <- dat_PW %>% 
  filter(intervention != "FollowUp") %>% 
  mutate(tpmAbd_log     = log(tpmAbd + 1),
         intervention   = case_when(treatment == "Baseline" ~ "Baseline",
                                    TRUE                    ~ intervention),
         age_centered50 = age - 50,
         bmi_centered25 = bmi_t0 - 25,
         age_cat        = case_when(age < 50 ~ paste0(min(age), "-49"),
                                    TRUE     ~ paste0("50-", max(age))),
         age_cat        = factor(age_cat),
         bmi_cat        = case_when(bmi_t0 < 25 ~ paste0("[", floor(min(bmi_t0)), ", 25)"),
                                    TRUE        ~ paste0("[25, ", ceiling(max(bmi_t0)), "]")),
         bmi_cat        = factor(bmi_cat),
         phase          = case_when(timepoint %in% c("T3","T4") ~ "phase 2",
                                    TRUE                        ~ "phase 1"),
         phase          = factor(phase),
         dailyFiber_cat  = case_when(dailyConsumption_fiber >= 30 ~ ">= 30g",
                                     TRUE                         ~ "< 30g"),
         dailyFiber_cat  = factor(dailyFiber_cat),
         baselineDiv_cat = case_when(baseline_div < median_baselineDiv ~ "[2.8, 3.85)",
                                     TRUE                              ~ "[3.85, 4.4)"),
         baselineDiv_cat = factor(baselineDiv_cat))

Limit pathway data to categories Metabolism, Genetic Information Processing and Cellular Processes.

Code

dat_PW <- dat_PW %>% 
  filter(taxonomy_level1 %in% c("Metabolism", "Genetic Information Processing", "Cellular Processes"))

Check how many genes / pathways were unmapped

Code

datList_relFun <- readAndPrepare_functionalData(path_KEGG_KO             = path_KEGGKO_funData,
                                                path_KEGG_pathways       = path_KEGGPathways_funData,
                                                path_sampleLookupXlsx    = path_sampleLookupXlsx,
                                                path_krautLookupXlsx     = path_krautLookupXlsx,
                                                abundance_unit           = "relAbd",
                                                exclude_sickParticipants = "taxonomy paper")

datCheck_KO <- datList_relFun$datKO_stoolSamples
datCheck_PW <- datList_relFun$datPathways_stoolSamples

avgUnmapped_KO <- datCheck_KO %>% 
  filter(KO == "unknown") %>% 
  select(-KO) %>% 
  unlist() %>% 
  mean()
relAbdMapped_PW <- datCheck_PW %>% 
  select(starts_with("sample_")) %>% 
  colSums()
avgUnmapped_PW <- mean(1 - relAbdMapped_PW)

Regarding the KO gene data, on average 75.6% of genes were unmapped over all samples.
Regarding the pathway data, on average 99.4% of pathways were unmapped over all samples.

Run screening

Run screening on 7123 genes and 164 pathways.

Code

lowAbundanceGenes <- dat_KO %>% 
  mutate(relevantAbundance = (tpmAbd >= 10)) %>% 
  group_by(FeatureID) %>% 
  dplyr::summarize(lowTPMabundance = (sum(relevantAbundance) < 0.1*n())) %>% 
  filter(lowTPMabundance) %>% 
  pull(FeatureID) %>% 
  as.character()

dat_KO <- dat_KO %>% 
  filter(!(FeatureID %in% lowAbundanceGenes))


lowAbundancePathways <- dat_PW %>% 
  mutate(relevantAbundance = (tpmAbd >= 10)) %>% 
  group_by(taxonomy_level3) %>% 
  dplyr::summarize(lowTPMabundance = (sum(relevantAbundance) < 0.1*n())) %>% 
  filter(lowTPMabundance) %>% 
  pull(taxonomy_level3) %>% 
  as.character()

dat_PW <- dat_PW %>% 
  filter(!(taxonomy_level3 %in% lowAbundancePathways))

But before, filtering out 4338 genes and 64 pathways with low TPM abundances (i.e., all that don’t have a minimum abundance of 10 TPM in at least 10% of all samples). Running the screening on the remaining 2785 genes and 100 pathways.

For each gene and pathway, a Generalized Mixed Regression Model is estimated, identical to the models estimated for the blood measurements for the blood paper, with response parameter the TPM abundance.

Model selection is performed by estimating either estimating (A) a Gaussian model, and (B) a Gaussian model on ‘log(y + 1)’ transformed values, or (A) a zero-inflated Gaussian model (with only an intercept predictor for the ZI part, as a general excess zero correction), and (B) a zero-inflated Gaussian model on ‘log(y + 1)’ transformed values for each gene and selecting the best performing one of the respective two as the final model. If at least 10% of a gene’s observations are zero values then the zero-inflated models are estimated, otherwise the non-zero-inflated models are estimated.

Notes:

• For the non-zero-inflated models the best model is chosen based on the share of explained deviance
• For the zero-inflated models the best model is chosen based on the lower average of all absolute residuals, since these models are estimated based on a quasi-likelihood approach and the deviance is not available.

Unused alternatives

• a zero-inflated Poisson model could have been estimated on rounded values. However, a Poisson distribution is not able to account for more strongly right-skewed distributions with a stronger probability mass at zero
• a Tweedie (p = 1.5) distribution would be a right-skewed continuous distribution with also potential probability mass at zero. However, since (1) it’s not a proper established distribution and (2) the used log transformation already accounts for right-skewedness we don’t need it and don’t use it

Correction for multiple testing is performed by applying the Benjamini-Hochberg (FDR) method to all interventions’ p-values.

Additionally, the intervention effects for every model are estimated in separate interactions with gender (male or female), age (above or below 50), baseline BMI (above or below 25), daily fiber consumption (above or below 30), baseline Shannon diversity (above or below tbd). Independently for every interaction variable (age, sex, BMI, fiber, diversity) a multiple testing correction is applied.

Note: For these screening analyses we always use a significance threshold of 0.1 instead of 0.05.

The models are estimated with mgcv::gam() and NBZIMM::lme.zig().

Code

# helper function to estimate the models
# - dat_model:     Dataset used for model estimation
# - model_type:    One of c("Gaussian", "ZI Gaussian")
# - model_formula: A model formula created with "as.formula"
estimate_model <- function(dat_model, model_type, model_formula) {
  
  checkmate::assert_data_frame(dat_model)
  checkmate::assert_choice(model_type, choices = c("Gaussian", "ZI Gaussian"))
  checkmate::assert_class(model_formula, classes = "formula")
  
  
  if (model_type == "Gaussian") {
    model <- gam(model_formula, method = "REML", family = "gaussian", data = dat_model)
    
  } else { # model_type = "ZI Gaussian"
    model <- tryCatch({ lme.zig(fixed     = model_formula,
                                zi_fixed  = ~ 1,
                                random    = ~ 1 | participant_id,
                                zi_random = NULL,
                                data      = dat_model) },
                      error = function(e) { return(NULL) })
  }
  
  return(model)
}

Code

# helper function to estimate the interaction models.
# Returns a list with all three estimated models and the respective three summary tables.
# Argument 'selected_model' is the label of the previously selected model type in the model selection process.
estimate_interactionModels <- function(dat_model, selected_model) {
  
  if (selected_model == "ZI Gaussian") {
    # age interaction model
    model_ageInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "ZI Gaussian",
                                   model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25 + intervention:age_cat"))
    if (is.null(model_ageInt)) { # if model estimation failed
      model_ageInt <- estimate_model(dat_model     = dat_model,
                                     model_type    = "Gaussian",
                                     model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:age_cat"))
    }
    # sex interaction model
    model_sexInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "ZI Gaussian",
                                   model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25 + intervention:gender"))
    if (is.null(model_sexInt)) { # if model estimation failed
      model_sexInt <- estimate_model(dat_model     = dat_model,
                                     model_type    = "Gaussian",
                                     model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:gender"))
    }
    # BMI interaction model
    model_bmiInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "ZI Gaussian",
                                   model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25 + intervention:bmi_cat"))
    if (is.null(model_bmiInt)) { # if model estimation failed
      model_bmiInt <- estimate_model(dat_model     = dat_model,
                                     model_type    = "Gaussian",
                                     model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:bmi_cat"))
    }
    # fiber interaction model
    model_fibInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "ZI Gaussian",
                                   model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25 + intervention:dailyFiber_cat"))
    if (is.null(model_fibInt)) { # if model estimation failed
      model_fibInt <- estimate_model(dat_model     = dat_model,
                                     model_type    = "Gaussian",
                                     model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:dailyFiber_cat"))
    }
    # baseline diversity interaction model
    model_divInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "ZI Gaussian",
                                   model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25 + intervention:baselineDiv_cat"))
    if (is.null(model_divInt)) { # if model estimation failed
      model_divInt <- estimate_model(dat_model     = dat_model,
                                     model_type    = "Gaussian",
                                     model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:baselineDiv_cat"))
    }
    
  } else if (selected_model == "ZI log-Gaussian") {
    # age interaction model
    model_ageInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "ZI Gaussian",
                                   model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25 + intervention:age_cat"))
    if (is.null(model_ageInt)) { # if model estimation failed
      model_ageInt <- estimate_model(dat_model     = dat_model,
                                     model_type    = "Gaussian",
                                     model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:age_cat"))
    }
    # sex interaction model
    model_sexInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "ZI Gaussian",
                                   model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25 + intervention:gender"))
    if (is.null(model_sexInt)) { # if model estimation failed
      model_sexInt <- estimate_model(dat_model     = dat_model,
                                     model_type    = "Gaussian",
                                     model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:gender"))
    }
    # BMI interaction model
    model_bmiInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "ZI Gaussian",
                                   model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25 + intervention:bmi_cat"))
    if (is.null(model_bmiInt)) { # if model estimation failed
      model_bmiInt <- estimate_model(dat_model     = dat_model,
                                     model_type    = "Gaussian",
                                     model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:bmi_cat"))
    }
    # fiber interaction model
    model_fibInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "ZI Gaussian",
                                   model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25 + intervention:dailyFiber_cat"))
    if (is.null(model_fibInt)) { # if model estimation failed
      model_fibInt <- estimate_model(dat_model     = dat_model,
                                     model_type    = "Gaussian",
                                     model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:dailyFiber_cat"))
    }
    # baseline diversity interaction model
    model_divInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "ZI Gaussian",
                                   model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25 + intervention:baselineDiv_cat"))
    if (is.null(model_divInt)) { # if model estimation failed
      model_divInt <- estimate_model(dat_model     = dat_model,
                                     model_type    = "Gaussian",
                                     model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:baselineDiv_cat"))
    }
    
  } else if (selected_model == "Gaussian") {
    # age interaction model
    model_ageInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "Gaussian",
                                   model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:age_cat"))
    # sex interaction model
    model_sexInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "Gaussian",
                                   model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:gender"))
    # BMI interaction model
    model_bmiInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "Gaussian",
                                   model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:bmi_cat"))
    # fiber interaction model
    model_fibInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "Gaussian",
                                   model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:dailyFiber_cat"))
    # baseline diversity interaction model
    model_divInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "Gaussian",
                                   model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:baselineDiv_cat"))
    
  } else { # model_label == "log-Gaussian"
    # age interaction model
    model_ageInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "Gaussian",
                                   model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:age_cat"))
    # sex interaction model
    model_sexInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "Gaussian",
                                   model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:gender"))
    # BMI interaction model
    model_bmiInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "Gaussian",
                                   model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:bmi_cat"))
    # fiber interaction model
    model_fibInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "Gaussian",
                                   model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:dailyFiber_cat"))
    # baseline diversity interaction model
    model_divInt <- estimate_model(dat_model     = dat_model,
                                   model_type    = "Gaussian",
                                   model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're') + intervention:baselineDiv_cat"))
  }
  
  
  ### prepare results
  # age interaction model
  if (class(model_ageInt)[1] == "zigmm") {
    tab_ageInt           <- summary(model_ageInt)$tTable
    colnames(tab_ageInt) <- c("Estimate", "Std. Error", "DF", "t-value", "Pr(>|t|)") # make names consistent to gam tab names
  } else { # non-ZI model
    tab_ageInt           <- summary(model_ageInt)$p.table
  }
  # sex interaction model
  if (class(model_sexInt)[1] == "zigmm") {
    tab_sexInt           <- summary(model_sexInt)$tTable
    colnames(tab_sexInt) <- c("Estimate", "Std. Error", "DF", "t-value", "Pr(>|t|)") # make names consistent to gam tab names
  } else { # non-ZI model
    tab_sexInt           <- summary(model_sexInt)$p.table
  }
  # BMI interaction model
  if (class(model_bmiInt)[1] == "zigmm") {
    tab_bmiInt           <- summary(model_bmiInt)$tTable
    colnames(tab_bmiInt) <- c("Estimate", "Std. Error", "DF", "t-value", "Pr(>|t|)") # make names consistent to gam tab names
  } else { # non-ZI model
    tab_bmiInt           <- summary(model_bmiInt)$p.table
  }
  # fiber interaction model
  if (class(model_fibInt)[1] == "zigmm") {
    tab_fibInt           <- summary(model_fibInt)$tTable
    colnames(tab_fibInt) <- c("Estimate", "Std. Error", "DF", "t-value", "Pr(>|t|)") # make names consistent to gam tab names
  } else { # non-ZI model
    tab_fibInt           <- summary(model_fibInt)$p.table
  }
  # baseline diversity interaction model
  if (class(model_divInt)[1] == "zigmm") {
    tab_divInt           <- summary(model_divInt)$tTable
    colnames(tab_divInt) <- c("Estimate", "Std. Error", "DF", "t-value", "Pr(>|t|)") # make names consistent to gam tab names
  } else { # non-ZI model
    tab_divInt           <- summary(model_divInt)$p.table
  }

  
  list(model_ageInt = model_ageInt,
       model_sexInt = model_sexInt,
       model_bmiInt = model_bmiInt,
       model_fibInt = model_fibInt,
       model_divInt = model_divInt,
       tab_ageInt   = tab_ageInt,
       tab_sexInt   = tab_sexInt,
       tab_bmiInt   = tab_bmiInt,
       tab_fibInt   = tab_fibInt,
       tab_divInt   = tab_divInt)
}

Code

# main screening function for genes
runScreening_oneGene <- function(gene) {
  
  message("Process gene ", gene, " (", match(gene, genes), "/", length(genes), ")")
  
  dat_gene <- dat_KO[dat_KO$FeatureID == gene,]
  
  # 1) main model estimation, based on model selection
  estimate_ZImodels <- sum(dat_gene$tpmAbd == 0) >= (0.1*nrow(dat_gene))
  if (estimate_ZImodels) { # estimate zero-inflated models
    
    # these zero-inflated models sometimes run into an estimation error. In this case, ignore the respective model.
    # If both models run into an estimation error, simply estimate the non-zero-inflated models instead.
    model_list <- list("ZI Gaussian"     = estimate_model(dat_model     = dat_gene,
                                                          model_type    = "ZI Gaussian",
                                                          model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25")),
                       "ZI log-Gaussian" = estimate_model(dat_model     = dat_gene,
                                                          model_type    = "ZI Gaussian",
                                                          model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25")))
    
    fallBack_toNonZImodels <- ifelse(is.null(model_list[[1]]) & is.null(model_list[[2]]), TRUE, FALSE)
    
    if (!fallBack_toNonZImodels) {
      
      if (is.null(model_list[[1]]) | is.null(model_list[[2]])) {
        null_index <- unname(which(sapply(model_list, function(x) { is.null(x) })))
        model_list <- model_list[-null_index]
      }
      
      bestModel_index <- unname(which.min(sapply(model_list, function(x) { mean(abs(summary(x)$residuals)) })))
      model           <- model_list[[bestModel_index]]
      model_label     <- names(model_list)[bestModel_index]
      tab             <- summary(model)$tTable
      colnames(tab)   <- c("Estimate", "Std. Error", "DF", "t-value", "Pr(>|t|)") # make names consistent to gam tab names
    }
    
  } else {
    fallBack_toNonZImodels <- FALSE # simply to have this object defined also if ZI models are not estimated
  }
  
  if (!estimate_ZImodels || fallBack_toNonZImodels) { # estimate models without zero-inflation
    
    model_list <- list("Gaussian"     = estimate_model(dat_model     = dat_gene,
                                                       model_type    = "Gaussian",
                                                       model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're')")),
                       "log-Gaussian" = estimate_model(dat_model     = dat_gene,
                                                       model_type    = "Gaussian",
                                                       model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're')")))
    
    bestModel_index <- unname(which.max(sapply(model_list, function(x) { summary(x)$dev.expl })))
    model           <- model_list[[bestModel_index]]
    model_label     <- names(model_list)[bestModel_index]
    tab             <- summary(model)$p.table
    
  }
  
  
  # 2) interaction model estimation, only for the final selected model
  # model estimation with the lower category of the interaction variable as the reference category
  model_intList <- estimate_interactionModels(dat_model      = dat_gene,
                                              selected_model = model_label)
  # model estimation with the higher category of the interaction variable as the reference category
  dat_gene_changedRef         <- dat_gene
  dat_gene_changedRef$gender  <- relevel(dat_gene_changedRef$gender,  ref = "W")
  dat_gene_changedRef$age_cat <- relevel(dat_gene_changedRef$age_cat, ref = "50-69")
  dat_gene_changedRef$bmi_cat <- relevel(dat_gene_changedRef$bmi_cat, ref = "[25, 31]")
  dat_gene_changedRef$dailyFiber_cat  <- relevel(dat_gene_changedRef$dailyFiber_cat,  ref = ">= 30g")
  dat_gene_changedRef$baselineDiv_cat <- relevel(dat_gene_changedRef$baselineDiv_cat, ref = "[3.85, 4.4)")
  model_intList_changedRef  <- estimate_interactionModels(dat_model      = dat_gene_changedRef,
                                                          selected_model = model_label)
  
  # create joint table
  tab_intEffects <- data.frame(effect_type    = c("fresh intervention", "pasteurized intervention"),
                               effect_ageLow  = c(model_intList$tab_ageInt["interventionFresh", "Estimate"],
                                                  model_intList$tab_ageInt["interventionPasteurized", "Estimate"]),
                               effect_ageHigh = c(model_intList_changedRef$tab_ageInt["interventionFresh", "Estimate"],
                                                  model_intList_changedRef$tab_ageInt["interventionPasteurized", "Estimate"]),
                               se_ageLow      = c(model_intList$tab_ageInt["interventionFresh", "Std. Error"],
                                                  model_intList$tab_ageInt["interventionPasteurized", "Std. Error"]),
                               se_ageHigh     = c(model_intList_changedRef$tab_ageInt["interventionFresh", "Std. Error"],
                                                  model_intList_changedRef$tab_ageInt["interventionPasteurized", "Std. Error"]),
                               pvalue_ageLow  = c(model_intList$tab_ageInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList$tab_ageInt["interventionPasteurized", "Pr(>|t|)"]),
                               pvalue_ageHigh = c(model_intList_changedRef$tab_ageInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList_changedRef$tab_ageInt["interventionPasteurized", "Pr(>|t|)"]),
                               effect_sexM    = c(model_intList$tab_sexInt["interventionFresh", "Estimate"],
                                                  model_intList$tab_sexInt["interventionPasteurized", "Estimate"]),
                               effect_sexW    = c(model_intList_changedRef$tab_sexInt["interventionFresh", "Estimate"],
                                                  model_intList_changedRef$tab_sexInt["interventionPasteurized", "Estimate"]),
                               se_sexM        = c(model_intList$tab_sexInt["interventionFresh", "Std. Error"],
                                                  model_intList$tab_sexInt["interventionPasteurized", "Std. Error"]),
                               se_sexW        = c(model_intList_changedRef$tab_sexInt["interventionFresh", "Std. Error"],
                                                  model_intList_changedRef$tab_sexInt["interventionPasteurized", "Std. Error"]),
                               pvalue_sexM    = c(model_intList$tab_sexInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList$tab_sexInt["interventionPasteurized", "Pr(>|t|)"]),
                               pvalue_sexW    = c(model_intList_changedRef$tab_sexInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList_changedRef$tab_sexInt["interventionPasteurized", "Pr(>|t|)"]),
                               effect_bmiLow  = c(model_intList$tab_bmiInt["interventionFresh", "Estimate"],
                                                  model_intList$tab_bmiInt["interventionPasteurized", "Estimate"]),
                               effect_bmiHigh = c(model_intList_changedRef$tab_bmiInt["interventionFresh", "Estimate"],
                                                  model_intList_changedRef$tab_bmiInt["interventionPasteurized", "Estimate"]),
                               se_bmiLow      = c(model_intList$tab_bmiInt["interventionFresh", "Std. Error"],
                                                  model_intList$tab_bmiInt["interventionPasteurized", "Std. Error"]),
                               se_bmiHigh     = c(model_intList_changedRef$tab_bmiInt["interventionFresh", "Std. Error"],
                                                  model_intList_changedRef$tab_bmiInt["interventionPasteurized", "Std. Error"]),
                               pvalue_bmiLow  = c(model_intList$tab_bmiInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList$tab_bmiInt["interventionPasteurized", "Pr(>|t|)"]),
                               pvalue_bmiHigh = c(model_intList_changedRef$tab_bmiInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList_changedRef$tab_bmiInt["interventionPasteurized", "Pr(>|t|)"]),
                               effect_fibLow  = c(model_intList$tab_fibInt["interventionFresh", "Estimate"],
                                                  model_intList$tab_fibInt["interventionPasteurized", "Estimate"]),
                               effect_fibHigh = c(model_intList_changedRef$tab_fibInt["interventionFresh", "Estimate"],
                                                  model_intList_changedRef$tab_fibInt["interventionPasteurized", "Estimate"]),
                               se_fibLow      = c(model_intList$tab_fibInt["interventionFresh", "Std. Error"],
                                                  model_intList$tab_fibInt["interventionPasteurized", "Std. Error"]),
                               se_fibHigh     = c(model_intList_changedRef$tab_fibInt["interventionFresh", "Std. Error"],
                                                  model_intList_changedRef$tab_fibInt["interventionPasteurized", "Std. Error"]),
                               pvalue_fibLow  = c(model_intList$tab_fibInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList$tab_fibInt["interventionPasteurized", "Pr(>|t|)"]),
                               pvalue_fibHigh = c(model_intList_changedRef$tab_fibInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList_changedRef$tab_fibInt["interventionPasteurized", "Pr(>|t|)"]),
                               effect_divLow  = c(model_intList$tab_divInt["interventionFresh", "Estimate"],
                                                  model_intList$tab_divInt["interventionPasteurized", "Estimate"]),
                               effect_divHigh = c(model_intList_changedRef$tab_divInt["interventionFresh", "Estimate"],
                                                  model_intList_changedRef$tab_divInt["interventionPasteurized", "Estimate"]),
                               se_divLow      = c(model_intList$tab_divInt["interventionFresh", "Std. Error"],
                                                  model_intList$tab_divInt["interventionPasteurized", "Std. Error"]),
                               se_divHigh     = c(model_intList_changedRef$tab_divInt["interventionFresh", "Std. Error"],
                                                  model_intList_changedRef$tab_divInt["interventionPasteurized", "Std. Error"]),
                               pvalue_divLow  = c(model_intList$tab_divInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList$tab_divInt["interventionPasteurized", "Pr(>|t|)"]),
                               pvalue_divHigh = c(model_intList_changedRef$tab_divInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList_changedRef$tab_divInt["interventionPasteurized", "Pr(>|t|)"]))
  
  
  # 3) gather results
  data.frame(gene               = gene,
             relFreq_zeroTPM    = sum(dat_gene$tpmAbd == 0) / nrow(dat_gene),
             relFreq_min10TPM   = sum(dat_gene$tpmAbd >= 10) / nrow(dat_gene),
             min_tpmAbd         = min(dat_gene$tpmAbd),
             mean_tpmAbd        = mean(dat_gene$tpmAbd),
             median_tpmAbd      = median(dat_gene$tpmAbd),
             max_tpmAbd         = max(dat_gene$tpmAbd),
             baseline_medianTPMAbd = median(dat_gene$tpmAbd[dat_gene$inter_treat %in% c("Baseline.Fresh","Baseline.Pasteurized")]),
             baseline_prevalence     = unname(prop.table(table(dat_gene$tpmAbd[dat_gene$inter_treat %in% c("Baseline.Fresh","Baseline.Pasteurized")] > 10))["TRUE"]),
             postFresh_medianTPMAbd  = median(dat_gene$tpmAbd[dat_gene$inter_treat == "After.Fresh"]),
             postFresh_prevalence    = unname(prop.table(table(dat_gene$tpmAbd[dat_gene$inter_treat == "After.Fresh"] > 10))["TRUE"]),
             postPast_medianTPMAbd   = median(dat_gene$tpmAbd[dat_gene$inter_treat == "After.Pasteurized"]),
             postPast_prevalence     = unname(prop.table(table(dat_gene$tpmAbd[dat_gene$inter_treat == "After.Pasteurized"] > 10))["TRUE"]),
             model              = model_label,
             model_deviance     = ifelse(!grepl("ZI", model_label), summary(model)$dev.expl, NA),
             model_meanAbsResid = ifelse(grepl("ZI", model_label), mean(abs(summary(model)$residuals)), NA),
             freshInt_effect    = tab["interventionFresh", "Estimate"],
             freshInt_se        = tab["interventionFresh", "Std. Error"],
             freshInt_pvalue    = tab["interventionFresh", "Pr(>|t|)"],
             pastInt_effect     = tab["interventionPasteurized", "Estimate"],
             pastInt_se         = tab["interventionPasteurized", "Std. Error"],
             pastInt_pvalue     = tab["interventionPasteurized", "Pr(>|t|)"],
             phaseEffect        = tab["phasephase 2", "Estimate"],
             genderEffect       = tab["genderW", "Estimate"],
             ageEffect          = tab["age_centered50", "Estimate"],
             bmiEffect          = tab["bmi_centered25", "Estimate"],
             freshInt_ageLow_effect  = tab_intEffects$effect_ageLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_ageHigh_effect = tab_intEffects$effect_ageHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_ageLow_se      = tab_intEffects$se_ageLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_ageHigh_se     = tab_intEffects$se_ageHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_ageLow_pvalue  = tab_intEffects$pvalue_ageLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_ageHigh_pvalue = tab_intEffects$pvalue_ageHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_sexM_effect    = tab_intEffects$effect_sexM[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_sexW_effect    = tab_intEffects$effect_sexW[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_sexM_se        = tab_intEffects$se_sexM[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_sexW_se        = tab_intEffects$se_sexW[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_sexM_pvalue    = tab_intEffects$pvalue_sexM[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_sexW_pvalue    = tab_intEffects$pvalue_sexW[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_bmiLow_effect  = tab_intEffects$effect_bmiLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_bmiHigh_effect = tab_intEffects$effect_bmiHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_bmiLow_se      = tab_intEffects$se_bmiLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_bmiHigh_se     = tab_intEffects$se_bmiHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_bmiLow_pvalue  = tab_intEffects$pvalue_bmiLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_bmiHigh_pvalue = tab_intEffects$pvalue_bmiHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_fibLow_effect  = tab_intEffects$effect_fibLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_fibHigh_effect = tab_intEffects$effect_fibHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_fibLow_se      = tab_intEffects$se_fibLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_fibHigh_se     = tab_intEffects$se_fibHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_fibLow_pvalue  = tab_intEffects$pvalue_fibLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_fibHigh_pvalue = tab_intEffects$pvalue_fibHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_divLow_effect  = tab_intEffects$effect_divLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_divHigh_effect = tab_intEffects$effect_divHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_divLow_se      = tab_intEffects$se_divLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_divHigh_se     = tab_intEffects$se_divHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_divLow_pvalue  = tab_intEffects$pvalue_divLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_divHigh_pvalue = tab_intEffects$pvalue_divHigh[tab_intEffects$effect_type == "fresh intervention"],
             pastInt_ageLow_effect   = tab_intEffects$effect_ageLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_ageHigh_effect  = tab_intEffects$effect_ageHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_ageLow_se       = tab_intEffects$se_ageLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_ageHigh_se      = tab_intEffects$se_ageHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_ageLow_pvalue   = tab_intEffects$pvalue_ageLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_ageHigh_pvalue  = tab_intEffects$pvalue_ageHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_sexM_effect     = tab_intEffects$effect_sexM[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_sexW_effect     = tab_intEffects$effect_sexW[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_sexM_se         = tab_intEffects$se_sexM[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_sexW_se         = tab_intEffects$se_sexW[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_sexM_pvalue     = tab_intEffects$pvalue_sexM[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_sexW_pvalue     = tab_intEffects$pvalue_sexW[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_bmiLow_effect   = tab_intEffects$effect_bmiLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_bmiHigh_effect  = tab_intEffects$effect_bmiHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_bmiLow_se       = tab_intEffects$se_bmiLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_bmiHigh_se      = tab_intEffects$se_bmiHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_bmiLow_pvalue   = tab_intEffects$pvalue_bmiLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_bmiHigh_pvalue  = tab_intEffects$pvalue_bmiHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_fibLow_effect   = tab_intEffects$effect_fibLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_fibHigh_effect  = tab_intEffects$effect_fibHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_fibLow_se       = tab_intEffects$se_fibLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_fibHigh_se      = tab_intEffects$se_fibHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_fibLow_pvalue   = tab_intEffects$pvalue_fibLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_fibHigh_pvalue  = tab_intEffects$pvalue_fibHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_divLow_effect   = tab_intEffects$effect_divLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_divHigh_effect  = tab_intEffects$effect_divHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_divLow_se       = tab_intEffects$se_divLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_divHigh_se      = tab_intEffects$se_divHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_divLow_pvalue   = tab_intEffects$pvalue_divLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_divHigh_pvalue  = tab_intEffects$pvalue_divHigh[tab_intEffects$effect_type == "pasteurized intervention"])
}


# main screening function for pathways
runScreening_onePathway <- function(pathway) {
  
  message("Process pathway ", pathway, " (", match(pathway, pathways), "/", length(pathways), ")")
  
  dat_pathway <- dat_PW[dat_PW$taxonomy_level3 == pathway,]
  
  # 1) main model estimation, based on model selection
  estimate_ZImodels <- sum(dat_pathway$tpmAbd == 0) >= (0.1*nrow(dat_pathway))
  if (estimate_ZImodels) { # estimate zero-inflated models
    
    # these zero-inflated models sometimes run into an estimation error. In this case, ignore the respective model.
    # If both models run into an estimation error, simply estimate the non-zero-inflated models instead.
    model_list <- list("ZI Gaussian"     = estimate_model(dat_model     = dat_pathway,
                                                          model_type    = "ZI Gaussian",
                                                          model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25")),
                       "ZI log-Gaussian" = estimate_model(dat_model     = dat_pathway,
                                                          model_type    = "ZI Gaussian",
                                                          model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25")))
    
    fallBack_toNonZImodels <- ifelse(is.null(model_list[[1]]) & is.null(model_list[[2]]), TRUE, FALSE)
    
    if (!fallBack_toNonZImodels) {
      
      if (is.null(model_list[[1]]) | is.null(model_list[[2]])) {
        null_index <- unname(which(sapply(model_list, function(x) { is.null(x) })))
        model_list <- model_list[-null_index]
      }
      
      bestModel_index <- unname(which.min(sapply(model_list, function(x) { mean(abs(summary(x)$residuals)) })))
      model           <- model_list[[bestModel_index]]
      model_label     <- names(model_list)[bestModel_index]
      tab             <- summary(model)$tTable
      colnames(tab)   <- c("Estimate", "Std. Error", "DF", "t-value", "Pr(>|t|)") # make names consistent to gam tab names
    }
    
  } else {
    fallBack_toNonZImodels <- FALSE # simply to have this object defined also if ZI models are not estimated
  }
  
  if (!estimate_ZImodels || fallBack_toNonZImodels) { # estimate models without zero-inflation
    
    model_list <- list("Gaussian"     = estimate_model(dat_model     = dat_pathway,
                                                       model_type    = "Gaussian",
                                                       model_formula = as.formula("tpmAbd ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're')")),
                       "log-Gaussian" = estimate_model(dat_model     = dat_pathway,
                                                       model_type    = "Gaussian",
                                                       model_formula = as.formula("tpmAbd_log ~ intervention + phase + gender + age_centered50 + bmi_centered25 + s(participant_id, bs = 're')")))
    
    bestModel_index <- unname(which.max(sapply(model_list, function(x) { summary(x)$dev.expl })))
    model           <- model_list[[bestModel_index]]
    model_label     <- names(model_list)[bestModel_index]
    tab             <- summary(model)$p.table
    
  }
  
  
  # 2) interaction model estimation, only for the final selected model
  # model estimation with the lower category of the interaction variable as the reference category
  model_intList <- estimate_interactionModels(dat_model      = dat_pathway,
                                              selected_model = model_label)
  # model estimation with the higher category of the interaction variable as the reference category
  dat_pathway_changedRef         <- dat_pathway
  dat_pathway_changedRef$gender  <- relevel(dat_pathway_changedRef$gender,  ref = "W")
  dat_pathway_changedRef$age_cat <- relevel(dat_pathway_changedRef$age_cat, ref = "50-69")
  dat_pathway_changedRef$bmi_cat <- relevel(dat_pathway_changedRef$bmi_cat, ref = "[25, 31]")
  dat_pathway_changedRef$dailyFiber_cat  <- relevel(dat_pathway_changedRef$dailyFiber_cat,  ref = ">= 30g")
  dat_pathway_changedRef$baselineDiv_cat <- relevel(dat_pathway_changedRef$baselineDiv_cat, ref = "[3.85, 4.4)")
  model_intList_changedRef  <- estimate_interactionModels(dat_model      = dat_pathway_changedRef,
                                                          selected_model = model_label)
  
  # create joint table
  tab_intEffects <- data.frame(effect_type    = c("fresh intervention", "pasteurized intervention"),
                               effect_ageLow  = c(model_intList$tab_ageInt["interventionFresh", "Estimate"],
                                                  model_intList$tab_ageInt["interventionPasteurized", "Estimate"]),
                               effect_ageHigh = c(model_intList_changedRef$tab_ageInt["interventionFresh", "Estimate"],
                                                  model_intList_changedRef$tab_ageInt["interventionPasteurized", "Estimate"]),
                               se_ageLow      = c(model_intList$tab_ageInt["interventionFresh", "Std. Error"],
                                                  model_intList$tab_ageInt["interventionPasteurized", "Std. Error"]),
                               se_ageHigh     = c(model_intList_changedRef$tab_ageInt["interventionFresh", "Std. Error"],
                                                  model_intList_changedRef$tab_ageInt["interventionPasteurized", "Std. Error"]),
                               pvalue_ageLow  = c(model_intList$tab_ageInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList$tab_ageInt["interventionPasteurized", "Pr(>|t|)"]),
                               pvalue_ageHigh = c(model_intList_changedRef$tab_ageInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList_changedRef$tab_ageInt["interventionPasteurized", "Pr(>|t|)"]),
                               effect_sexM    = c(model_intList$tab_sexInt["interventionFresh", "Estimate"],
                                                  model_intList$tab_sexInt["interventionPasteurized", "Estimate"]),
                               effect_sexW    = c(model_intList_changedRef$tab_sexInt["interventionFresh", "Estimate"],
                                                  model_intList_changedRef$tab_sexInt["interventionPasteurized", "Estimate"]),
                               se_sexM        = c(model_intList$tab_sexInt["interventionFresh", "Std. Error"],
                                                  model_intList$tab_sexInt["interventionPasteurized", "Std. Error"]),
                               se_sexW        = c(model_intList_changedRef$tab_sexInt["interventionFresh", "Std. Error"],
                                                  model_intList_changedRef$tab_sexInt["interventionPasteurized", "Std. Error"]),
                               pvalue_sexM    = c(model_intList$tab_sexInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList$tab_sexInt["interventionPasteurized", "Pr(>|t|)"]),
                               pvalue_sexW    = c(model_intList_changedRef$tab_sexInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList_changedRef$tab_sexInt["interventionPasteurized", "Pr(>|t|)"]),
                               effect_bmiLow  = c(model_intList$tab_bmiInt["interventionFresh", "Estimate"],
                                                  model_intList$tab_bmiInt["interventionPasteurized", "Estimate"]),
                               effect_bmiHigh = c(model_intList_changedRef$tab_bmiInt["interventionFresh", "Estimate"],
                                                  model_intList_changedRef$tab_bmiInt["interventionPasteurized", "Estimate"]),
                               se_bmiLow      = c(model_intList$tab_bmiInt["interventionFresh", "Std. Error"],
                                                  model_intList$tab_bmiInt["interventionPasteurized", "Std. Error"]),
                               se_bmiHigh     = c(model_intList_changedRef$tab_bmiInt["interventionFresh", "Std. Error"],
                                                  model_intList_changedRef$tab_bmiInt["interventionPasteurized", "Std. Error"]),
                               pvalue_bmiLow  = c(model_intList$tab_bmiInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList$tab_bmiInt["interventionPasteurized", "Pr(>|t|)"]),
                               pvalue_bmiHigh = c(model_intList_changedRef$tab_bmiInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList_changedRef$tab_bmiInt["interventionPasteurized", "Pr(>|t|)"]),
                               effect_fibLow  = c(model_intList$tab_fibInt["interventionFresh", "Estimate"],
                                                  model_intList$tab_fibInt["interventionPasteurized", "Estimate"]),
                               effect_fibHigh = c(model_intList_changedRef$tab_fibInt["interventionFresh", "Estimate"],
                                                  model_intList_changedRef$tab_fibInt["interventionPasteurized", "Estimate"]),
                               se_fibLow      = c(model_intList$tab_fibInt["interventionFresh", "Std. Error"],
                                                  model_intList$tab_fibInt["interventionPasteurized", "Std. Error"]),
                               se_fibHigh     = c(model_intList_changedRef$tab_fibInt["interventionFresh", "Std. Error"],
                                                  model_intList_changedRef$tab_fibInt["interventionPasteurized", "Std. Error"]),
                               pvalue_fibLow  = c(model_intList$tab_fibInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList$tab_fibInt["interventionPasteurized", "Pr(>|t|)"]),
                               pvalue_fibHigh = c(model_intList_changedRef$tab_fibInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList_changedRef$tab_fibInt["interventionPasteurized", "Pr(>|t|)"]),
                               effect_divLow  = c(model_intList$tab_divInt["interventionFresh", "Estimate"],
                                                  model_intList$tab_divInt["interventionPasteurized", "Estimate"]),
                               effect_divHigh = c(model_intList_changedRef$tab_divInt["interventionFresh", "Estimate"],
                                                  model_intList_changedRef$tab_divInt["interventionPasteurized", "Estimate"]),
                               se_divLow      = c(model_intList$tab_divInt["interventionFresh", "Std. Error"],
                                                  model_intList$tab_divInt["interventionPasteurized", "Std. Error"]),
                               se_divHigh     = c(model_intList_changedRef$tab_divInt["interventionFresh", "Std. Error"],
                                                  model_intList_changedRef$tab_divInt["interventionPasteurized", "Std. Error"]),
                               pvalue_divLow  = c(model_intList$tab_divInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList$tab_divInt["interventionPasteurized", "Pr(>|t|)"]),
                               pvalue_divHigh = c(model_intList_changedRef$tab_divInt["interventionFresh", "Pr(>|t|)"],
                                                  model_intList_changedRef$tab_divInt["interventionPasteurized", "Pr(>|t|)"]))
  
  
  # 3) gather results
  data.frame(pathway            = pathway,
             relFreq_zeroTPM    = sum(dat_pathway$tpmAbd == 0) / nrow(dat_pathway),
             relFreq_min10TPM   = sum(dat_pathway$tpmAbd >= 10) / nrow(dat_pathway),
             min_tpmAbd         = min(dat_pathway$tpmAbd),
             mean_tpmAbd        = mean(dat_pathway$tpmAbd),
             median_tpmAbd      = median(dat_pathway$tpmAbd),
             max_tpmAbd         = max(dat_pathway$tpmAbd),
             baseline_medianTPMAbd = median(dat_pathway$tpmAbd[dat_pathway$inter_treat %in% c("Baseline.Fresh","Baseline.Pasteurized")]),
             baseline_prevalence     = unname(prop.table(table(dat_pathway$tpmAbd[dat_pathway$inter_treat %in% c("Baseline.Fresh","Baseline.Pasteurized")] > 10))["TRUE"]),
             postFresh_medianTPMAbd  = median(dat_pathway$tpmAbd[dat_pathway$inter_treat == "After.Fresh"]),
             postFresh_prevalence    = unname(prop.table(table(dat_pathway$tpmAbd[dat_pathway$inter_treat == "After.Fresh"] > 10))["TRUE"]),
             postPast_medianTPMAbd   = median(dat_pathway$tpmAbd[dat_pathway$inter_treat == "After.Pasteurized"]),
             postPast_prevalence     = unname(prop.table(table(dat_pathway$tpmAbd[dat_pathway$inter_treat == "After.Pasteurized"] > 10))["TRUE"]),
             model              = model_label,
             model_deviance     = ifelse(!grepl("ZI", model_label), summary(model)$dev.expl, NA),
             model_meanAbsResid = ifelse(grepl("ZI", model_label), mean(abs(summary(model)$residuals)), NA),
             freshInt_effect    = tab["interventionFresh", "Estimate"],
             freshInt_se        = tab["interventionFresh", "Std. Error"],
             freshInt_pvalue    = tab["interventionFresh", "Pr(>|t|)"],
             pastInt_effect     = tab["interventionPasteurized", "Estimate"],
             pastInt_se         = tab["interventionPasteurized", "Std. Error"],
             pastInt_pvalue     = tab["interventionPasteurized", "Pr(>|t|)"],
             phaseEffect        = tab["phasephase 2", "Estimate"],
             genderEffect       = tab["genderW", "Estimate"],
             ageEffect          = tab["age_centered50", "Estimate"],
             bmiEffect          = tab["bmi_centered25", "Estimate"],
             freshInt_ageLow_effect  = tab_intEffects$effect_ageLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_ageHigh_effect = tab_intEffects$effect_ageHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_ageLow_se      = tab_intEffects$se_ageLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_ageHigh_se     = tab_intEffects$se_ageHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_ageLow_pvalue  = tab_intEffects$pvalue_ageLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_ageHigh_pvalue = tab_intEffects$pvalue_ageHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_sexM_effect    = tab_intEffects$effect_sexM[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_sexW_effect    = tab_intEffects$effect_sexW[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_sexM_se        = tab_intEffects$se_sexM[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_sexW_se        = tab_intEffects$se_sexW[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_sexM_pvalue    = tab_intEffects$pvalue_sexM[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_sexW_pvalue    = tab_intEffects$pvalue_sexW[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_bmiLow_effect  = tab_intEffects$effect_bmiLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_bmiHigh_effect = tab_intEffects$effect_bmiHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_bmiLow_se      = tab_intEffects$se_bmiLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_bmiHigh_se     = tab_intEffects$se_bmiHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_bmiLow_pvalue  = tab_intEffects$pvalue_bmiLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_bmiHigh_pvalue = tab_intEffects$pvalue_bmiHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_fibLow_effect  = tab_intEffects$effect_fibLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_fibHigh_effect = tab_intEffects$effect_fibHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_fibLow_se      = tab_intEffects$se_fibLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_fibHigh_se     = tab_intEffects$se_fibHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_fibLow_pvalue  = tab_intEffects$pvalue_fibLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_fibHigh_pvalue = tab_intEffects$pvalue_fibHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_divLow_effect  = tab_intEffects$effect_divLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_divHigh_effect = tab_intEffects$effect_divHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_divLow_se      = tab_intEffects$se_divLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_divHigh_se     = tab_intEffects$se_divHigh[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_divLow_pvalue  = tab_intEffects$pvalue_divLow[tab_intEffects$effect_type == "fresh intervention"],
             freshInt_divHigh_pvalue = tab_intEffects$pvalue_divHigh[tab_intEffects$effect_type == "fresh intervention"],
             pastInt_ageLow_effect   = tab_intEffects$effect_ageLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_ageHigh_effect  = tab_intEffects$effect_ageHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_ageLow_se       = tab_intEffects$se_ageLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_ageHigh_se      = tab_intEffects$se_ageHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_ageLow_pvalue   = tab_intEffects$pvalue_ageLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_ageHigh_pvalue  = tab_intEffects$pvalue_ageHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_sexM_effect     = tab_intEffects$effect_sexM[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_sexW_effect     = tab_intEffects$effect_sexW[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_sexM_se         = tab_intEffects$se_sexM[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_sexW_se         = tab_intEffects$se_sexW[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_sexM_pvalue     = tab_intEffects$pvalue_sexM[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_sexW_pvalue     = tab_intEffects$pvalue_sexW[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_bmiLow_effect   = tab_intEffects$effect_bmiLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_bmiHigh_effect  = tab_intEffects$effect_bmiHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_bmiLow_se       = tab_intEffects$se_bmiLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_bmiHigh_se      = tab_intEffects$se_bmiHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_bmiLow_pvalue   = tab_intEffects$pvalue_bmiLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_bmiHigh_pvalue  = tab_intEffects$pvalue_bmiHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_fibLow_effect   = tab_intEffects$effect_fibLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_fibHigh_effect  = tab_intEffects$effect_fibHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_fibLow_se       = tab_intEffects$se_fibLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_fibHigh_se      = tab_intEffects$se_fibHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_fibLow_pvalue   = tab_intEffects$pvalue_fibLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_fibHigh_pvalue  = tab_intEffects$pvalue_fibHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_divLow_effect   = tab_intEffects$effect_divLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_divHigh_effect  = tab_intEffects$effect_divHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_divLow_se       = tab_intEffects$se_divLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_divHigh_se      = tab_intEffects$se_divHigh[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_divLow_pvalue   = tab_intEffects$pvalue_divLow[tab_intEffects$effect_type == "pasteurized intervention"],
             pastInt_divHigh_pvalue  = tab_intEffects$pvalue_divHigh[tab_intEffects$effect_type == "pasteurized intervention"])
}

Code

# code for running the screening. It takes about 25 minutes on 15 cores
# to estimate the results. Instead of newly estimating them each time
# the html is generated, the results were saved after running the screening
# once, and are now just read from the results csv
dat_KOresults <- read.csv("9_genesAndPathways_regressionScreening_KOresults.csv")

# genes <- dat_KO %>% filter(FeatureID != "unknown") %>% pull(FeatureID) %>% unique() %>% as.character()
# 
# # number of cores to use for the parallelized call
# n_cores <- 15
# 
# # create a cluster object for Windows parallelization
# cl <- makeCluster(n_cores)
# 
# # export necessary functions / data objects to the cluster workers
# clusterExport(cl, c("runScreening_oneGene", "estimate_model", "estimate_interactionModels",
#                     "genes", "dat_KO", "gam", "lme.zig", "summary.gam"))
# 
# # run the screening (parallelized call)
# datList_KOresults <- parLapply(cl, genes, runScreening_oneGene)
# 
# # stop the Windows cluster
# stopCluster(cl)
# 
# # merge results datasets
# dat_KOresults <- datList_KOresults %>% dplyr::bind_rows()
# 
# # perform multiple testing correction and round values in the data
# p_corrected       <- p.adjust(c(dat_KOresults$freshInt_pvalue, dat_KOresults$pastInt_pvalue), method = "BH")
# pAgeInt_corrected <- p.adjust(c(dat_KOresults$freshInt_ageLow_pvalue,  dat_KOresults$pastInt_ageLow_pvalue,
#                                 dat_KOresults$freshInt_ageHigh_pvalue, dat_KOresults$pastInt_ageHigh_pvalue), method = "BH")
# pSexInt_corrected <- p.adjust(c(dat_KOresults$freshInt_sexM_pvalue,  dat_KOresults$pastInt_sexM_pvalue,
#                                 dat_KOresults$freshInt_sexW_pvalue, dat_KOresults$pastInt_sexW_pvalue), method = "BH")
# pBMIInt_corrected <- p.adjust(c(dat_KOresults$freshInt_bmiLow_pvalue,  dat_KOresults$pastInt_bmiLow_pvalue,
#                                 dat_KOresults$freshInt_bmiHigh_pvalue, dat_KOresults$pastInt_bmiHigh_pvalue), method = "BH")
# pFibInt_corrected <- p.adjust(c(dat_KOresults$freshInt_fibLow_pvalue,  dat_KOresults$pastInt_fibLow_pvalue,
#                                 dat_KOresults$freshInt_fibHigh_pvalue, dat_KOresults$pastInt_fibHigh_pvalue), method = "BH")
# pDivInt_corrected <- p.adjust(c(dat_KOresults$freshInt_divLow_pvalue,  dat_KOresults$pastInt_divLow_pvalue,
#                                 dat_KOresults$freshInt_divHigh_pvalue, dat_KOresults$pastInt_divHigh_pvalue), method = "BH")
# n             <- nrow(dat_KOresults)
# dat_KOresults <- dat_KOresults %>%
#   mutate(freshInt_pvalue_correct          = p_corrected[1:n],
#          pastInt_pvalue_correct           = p_corrected[(n+1):(2*n)],
#          freshInt_ageLow_pvalue_correct   = pAgeInt_corrected[1:n],
#          pastInt_ageLow_pvalue_correct    = pAgeInt_corrected[(n+1):(2*n)],
#          freshInt_ageHigh_pvalue_correct  = pAgeInt_corrected[(2*n+1):(3*n)],
#          pastInt_ageHigh_pvalue_correct   = pAgeInt_corrected[(3*n+1):(4*n)],
#          freshInt_sexM_pvalue_correct     = pSexInt_corrected[1:n],
#          pastInt_sexM_pvalue_correct      = pSexInt_corrected[(n+1):(2*n)],
#          freshInt_sexW_pvalue_correct     = pSexInt_corrected[(2*n+1):(3*n)],
#          pastInt_sexW_pvalue_correct      = pSexInt_corrected[(3*n+1):(4*n)],
#          freshInt_bmiLow_pvalue_correct   = pBMIInt_corrected[1:n],
#          pastInt_bmiLow_pvalue_correct    = pBMIInt_corrected[(n+1):(2*n)],
#          freshInt_bmiHigh_pvalue_correct  = pBMIInt_corrected[(2*n+1):(3*n)],
#          pastInt_bmiHigh_pvalue_correct   = pBMIInt_corrected[(3*n+1):(4*n)],
#          freshInt_fibLow_pvalue_correct   = pFibInt_corrected[1:n],
#          pastInt_fibLow_pvalue_correct    = pFibInt_corrected[(n+1):(2*n)],
#          freshInt_fibHigh_pvalue_correct  = pFibInt_corrected[(2*n+1):(3*n)],
#          pastInt_fibHigh_pvalue_correct   = pFibInt_corrected[(3*n+1):(4*n)],
#          freshInt_divLow_pvalue_correct   = pDivInt_corrected[1:n],
#          pastInt_divLow_pvalue_correct    = pDivInt_corrected[(n+1):(2*n)],
#          freshInt_divHigh_pvalue_correct  = pDivInt_corrected[(2*n+1):(3*n)],
#          pastInt_divHigh_pvalue_correct   = pDivInt_corrected[(3*n+1):(4*n)]) %>%
#   mutate_at(c(2:13, 15:ncol(.)), function(x) { round(x, 4) })
# 
# # add information if the genes relate to specific SCFAs
# dat_KOresults <- dat_KOresults %>%
#   dplyr::left_join(dat_scfaLookup, by = c("gene" = "KO")) %>%
#   dplyr::select(SCFA, everything())
# 
# write.csv(dat_KOresults, file = "9_genesAndPathways_regressionScreening_KOresults.csv", row.names = FALSE)

Code

# code for running the screening. It takes ~30 seconds on 15 cores
# to estimate the results. Instead of newly estimating them each time
# the html is generated, the results were saved after running the screening
# once, and are now just read from the results csv
dat_PWresults <- read.csv("9_genesAndPathways_regressionScreening_PWresults.csv")

# pathways <- dat_PW %>% pull(taxonomy_level3) %>% unique() %>% as.character()
# 
# # number of cores to use for the parallelized call
# n_cores <- 15
# 
# # create a cluster object for Windows parallelization
# cl <- makeCluster(n_cores)
# 
# # export necessary functions / data objects to the cluster workers
# clusterExport(cl, c("runScreening_onePathway", "estimate_model", "estimate_interactionModels",
#                     "pathways", "dat_PW", "gam", "lme.zig", "summary.gam"))
# 
# # run the screening (parallelized call)
# datList_PWresults <- parLapply(cl, pathways, runScreening_onePathway)
# 
# # stop the Windows cluster
# stopCluster(cl)
# 
# # merge results datasets
# dat_PWresults <- datList_PWresults %>% dplyr::bind_rows()
# 
# # perform multiple testing correction and round values in the data
# p_corrected       <- p.adjust(c(dat_PWresults$freshInt_pvalue, dat_PWresults$pastInt_pvalue), method = "BH")
# pAgeInt_corrected <- p.adjust(c(dat_PWresults$freshInt_ageLow_pvalue,  dat_PWresults$pastInt_ageLow_pvalue,
#                                 dat_PWresults$freshInt_ageHigh_pvalue, dat_PWresults$pastInt_ageHigh_pvalue), method = "BH")
# pSexInt_corrected <- p.adjust(c(dat_PWresults$freshInt_sexM_pvalue,  dat_PWresults$pastInt_sexM_pvalue,
#                                 dat_PWresults$freshInt_sexW_pvalue, dat_PWresults$pastInt_sexW_pvalue), method = "BH")
# pBMIInt_corrected <- p.adjust(c(dat_PWresults$freshInt_bmiLow_pvalue,  dat_PWresults$pastInt_bmiLow_pvalue,
#                                 dat_PWresults$freshInt_bmiHigh_pvalue, dat_PWresults$pastInt_bmiHigh_pvalue), method = "BH")
# pFibInt_corrected <- p.adjust(c(dat_PWresults$freshInt_fibLow_pvalue,  dat_PWresults$pastInt_fibLow_pvalue,
#                                 dat_PWresults$freshInt_fibHigh_pvalue, dat_PWresults$pastInt_fibHigh_pvalue), method = "BH")
# pDivInt_corrected <- p.adjust(c(dat_PWresults$freshInt_divLow_pvalue,  dat_PWresults$pastInt_divLow_pvalue,
#                                 dat_PWresults$freshInt_divHigh_pvalue, dat_PWresults$pastInt_divHigh_pvalue), method = "BH")
# n             <- nrow(dat_PWresults)
# dat_PWresults <- dat_PWresults %>%
#   mutate(freshInt_pvalue_correct          = p_corrected[1:n],
#          pastInt_pvalue_correct           = p_corrected[(n+1):(2*n)],
#          freshInt_ageLow_pvalue_correct   = pAgeInt_corrected[1:n],
#          pastInt_ageLow_pvalue_correct    = pAgeInt_corrected[(n+1):(2*n)],
#          freshInt_ageHigh_pvalue_correct  = pAgeInt_corrected[(2*n+1):(3*n)],
#          pastInt_ageHigh_pvalue_correct   = pAgeInt_corrected[(3*n+1):(4*n)],
#          freshInt_sexM_pvalue_correct     = pSexInt_corrected[1:n],
#          pastInt_sexM_pvalue_correct      = pSexInt_corrected[(n+1):(2*n)],
#          freshInt_sexW_pvalue_correct     = pSexInt_corrected[(2*n+1):(3*n)],
#          pastInt_sexW_pvalue_correct      = pSexInt_corrected[(3*n+1):(4*n)],
#          freshInt_bmiLow_pvalue_correct   = pBMIInt_corrected[1:n],
#          pastInt_bmiLow_pvalue_correct    = pBMIInt_corrected[(n+1):(2*n)],
#          freshInt_bmiHigh_pvalue_correct  = pBMIInt_corrected[(2*n+1):(3*n)],
#          pastInt_bmiHigh_pvalue_correct   = pBMIInt_corrected[(3*n+1):(4*n)],
#          freshInt_fibLow_pvalue_correct   = pFibInt_corrected[1:n],
#          pastInt_fibLow_pvalue_correct    = pFibInt_corrected[(n+1):(2*n)],
#          freshInt_fibHigh_pvalue_correct  = pFibInt_corrected[(2*n+1):(3*n)],
#          pastInt_fibHigh_pvalue_correct   = pFibInt_corrected[(3*n+1):(4*n)],
#          freshInt_divLow_pvalue_correct   = pDivInt_corrected[1:n],
#          pastInt_divLow_pvalue_correct    = pDivInt_corrected[(n+1):(2*n)],
#          freshInt_divHigh_pvalue_correct  = pDivInt_corrected[(2*n+1):(3*n)],
#          pastInt_divHigh_pvalue_correct   = pDivInt_corrected[(3*n+1):(4*n)]) %>%
#   mutate_at(c(2:13, 15:ncol(.)), function(x) { round(x, 4) })
# 
# write.csv(dat_PWresults, file = "9_genesAndPathways_regressionScreening_PWresults.csv", row.names = FALSE)

SCFA genes

Code

dat_SCFAresults <- dat_KOresults %>% filter(!is.na(SCFA))

Perform multiple testing correction of p-values only on the SCFA tests

Currently, this doesn’t change anything in the number of significant genes. Re-checked this, and everything is already. That’s just the result.

Code

p_corrected       <- p.adjust(c(dat_SCFAresults$freshInt_pvalue, dat_SCFAresults$pastInt_pvalue), method = "BH")
pAgeInt_corrected <- p.adjust(c(dat_SCFAresults$freshInt_ageLow_pvalue,  dat_SCFAresults$pastInt_ageLow_pvalue,
                                dat_SCFAresults$freshInt_ageHigh_pvalue, dat_SCFAresults$pastInt_ageHigh_pvalue), method = "BH")
pSexInt_corrected <- p.adjust(c(dat_SCFAresults$freshInt_sexM_pvalue,  dat_SCFAresults$pastInt_sexM_pvalue,
                                dat_SCFAresults$freshInt_sexW_pvalue, dat_SCFAresults$pastInt_sexW_pvalue), method = "BH")
pBMIInt_corrected <- p.adjust(c(dat_SCFAresults$freshInt_bmiLow_pvalue,  dat_SCFAresults$pastInt_bmiLow_pvalue,
                                dat_SCFAresults$freshInt_bmiHigh_pvalue, dat_SCFAresults$pastInt_bmiHigh_pvalue), method = "BH")
pFibInt_corrected <- p.adjust(c(dat_SCFAresults$freshInt_fibLow_pvalue,  dat_SCFAresults$pastInt_fibLow_pvalue,
                                dat_SCFAresults$freshInt_fibHigh_pvalue, dat_SCFAresults$pastInt_fibHigh_pvalue), method = "BH")
pDivInt_corrected <- p.adjust(c(dat_SCFAresults$freshInt_divLow_pvalue,  dat_SCFAresults$pastInt_divLow_pvalue,
                                dat_SCFAresults$freshInt_divHigh_pvalue, dat_SCFAresults$pastInt_divHigh_pvalue), method = "BH")
n               <- nrow(dat_SCFAresults)
dat_SCFAresults <- dat_SCFAresults %>%
  mutate(freshInt_pvalue_correct          = p_corrected[1:n] %>% round(4),
         pastInt_pvalue_correct           = p_corrected[(n+1):(2*n)] %>% round(4),
         freshInt_ageLow_pvalue_correct   = pAgeInt_corrected[1:n] %>% round(4),
         pastInt_ageLow_pvalue_correct    = pAgeInt_corrected[(n+1):(2*n)] %>% round(4),
         freshInt_ageHigh_pvalue_correct  = pAgeInt_corrected[(2*n+1):(3*n)] %>% round(4),
         pastInt_ageHigh_pvalue_correct   = pAgeInt_corrected[(3*n+1):(4*n)] %>% round(4),
         freshInt_sexM_pvalue_correct     = pSexInt_corrected[1:n] %>% round(4),
         pastInt_sexM_pvalue_correct      = pSexInt_corrected[(n+1):(2*n)] %>% round(4),
         freshInt_sexW_pvalue_correct     = pSexInt_corrected[(2*n+1):(3*n)] %>% round(4),
         pastInt_sexW_pvalue_correct      = pSexInt_corrected[(3*n+1):(4*n)] %>% round(4),
         freshInt_bmiLow_pvalue_correct   = pBMIInt_corrected[1:n] %>% round(4),
         pastInt_bmiLow_pvalue_correct    = pBMIInt_corrected[(n+1):(2*n)] %>% round(4),
         freshInt_bmiHigh_pvalue_correct  = pBMIInt_corrected[(2*n+1):(3*n)] %>% round(4),
         pastInt_bmiHigh_pvalue_correct   = pBMIInt_corrected[(3*n+1):(4*n)] %>% round(4),
         freshInt_fibLow_pvalue_correct   = pFibInt_corrected[1:n] %>% round(4),
         pastInt_fibLow_pvalue_correct    = pFibInt_corrected[(n+1):(2*n)] %>% round(4),
         freshInt_fibHigh_pvalue_correct  = pFibInt_corrected[(2*n+1):(3*n)] %>% round(4),
         pastInt_fibHigh_pvalue_correct   = pFibInt_corrected[(3*n+1):(4*n)] %>% round(4),
         freshInt_divLow_pvalue_correct   = pDivInt_corrected[1:n] %>% round(4),
         pastInt_divLow_pvalue_correct    = pDivInt_corrected[(n+1):(2*n)] %>% round(4),
         freshInt_divHigh_pvalue_correct  = pDivInt_corrected[(2*n+1):(3*n)] %>% round(4),
         pastInt_divHigh_pvalue_correct   = pDivInt_corrected[(3*n+1):(4*n)] %>% round(4)) %>% 
  mutate_at(c(3:14, 16:ncol(.)), function(x) { round(x, 4) })

Code

# some description
# dat_KO %>% 
#   dplyr::left_join(dat_scfaLookup, by = c("FeatureID" = "KO")) %>% 
#   filter(!is.na(SCFA),
#          intervention != "FollowUp") %>% 
#   group_by(FeatureID, timepoint) %>% 
#   dplyr::summarize(sampleShare_atLeast100TPM = sum(tpmAbd >= 100) / n(),
#             inter_treat               = first(inter_treat)) %>% 
#   group_by(FeatureID) %>% 
#   mutate(minY_forSorting = min(sampleShare_atLeast100TPM)) %>% 
#   ungroup() %>%
#   arrange(desc(minY_forSorting)) %>% 
#   mutate(FeatureID = factor(FeatureID, levels = unique(as.character(FeatureID)))) %>% 
#   ggplot(aes(y = FeatureID, x = inter_treat, fill = sampleShare_atLeast100TPM)) +
#   geom_raster()

SCFA results

Overview of the numbers of significant genes

Code

data.frame("what"  = c("No. of SCFA genes",
                       "... that didn't have low abundance",
                       "No. of significant genes (fresh intervention)",
                       "No. of significant genes (past. intervention)"),
           "value" = c(nrow(dat_scfaLookup),
                       nrow(dat_SCFAresults),
                       sum(dat_SCFAresults$freshInt_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_pvalue_correct < 0.1, na.rm = TRUE))) %>% 
  kable() %>% 
  kable_styling()

what	value
No. of SCFA genes	193
... that didn't have low abundance	89
No. of significant genes (fresh intervention)	0
No. of significant genes (past. intervention)	2

… alternatively, looking at the raw p-values, not corrected for multiple testing

Code

data.frame("what"  = c("No. of SCFA genes",
                       "... that didn't have low abundance",
                       "No. of significant genes (fresh intervention)",
                       "No. of significant genes (past. intervention)"),
           "value" = c(nrow(dat_scfaLookup),
                       nrow(dat_SCFAresults),
                       sum(dat_SCFAresults$freshInt_pvalue < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_pvalue < 0.1, na.rm = TRUE))) %>% 
  kable() %>% 
  kable_styling()

what	value
No. of SCFA genes	193
... that didn't have low abundance	89
No. of significant genes (fresh intervention)	4
No. of significant genes (past. intervention)	26

Overview which specific genes are significant (without multiple testing correction)

Code

dat_SCFAresults %>% 
  filter((freshInt_pvalue < 0.1) |
           (pastInt_pvalue < 0.1)) %>% 
  dplyr::select(-contains("correct")) %>% 
  mutate(across(contains("pvalue"), function(x) { ifelse(x < 0.1, x, "not significant") })) %>% 
  dplyr::select(SCFA, gene, contains("pvalue")) %>% 
  arrange(freshInt_pvalue, pastInt_pvalue) %>% 
  kable() %>% 
  kable_styling()

SCFA	gene	freshInt_pvalue	pastInt_pvalue	freshInt_ageLow_pvalue	freshInt_ageHigh_pvalue	freshInt_sexM_pvalue	freshInt_sexW_pvalue	freshInt_bmiLow_pvalue	freshInt_bmiHigh_pvalue	freshInt_fibLow_pvalue	freshInt_fibHigh_pvalue	freshInt_divLow_pvalue	freshInt_divHigh_pvalue	pastInt_ageLow_pvalue	pastInt_ageHigh_pvalue	pastInt_sexM_pvalue	pastInt_sexW_pvalue	pastInt_bmiLow_pvalue	pastInt_bmiHigh_pvalue	pastInt_fibLow_pvalue	pastInt_fibHigh_pvalue	pastInt_divLow_pvalue	pastInt_divHigh_pvalue
Pyruvate	K13290	0.0248	not significant	not significant	0.0673	not significant	0.0911	not significant	0.027	0.0236	not significant	0.0903	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant
Pyruvate	K13497	0.0297	0	0.0373	not significant	not significant	0.0395	0.0162	not significant	not significant	0.0416	not significant	not significant	0	not significant	0.0613	0.0001	0.0001	not significant	0.0001	0.0746	0.0132	0.0006
Pyruvate	K01573	0.0783	not significant	not significant	0.0168	0.046	not significant	not significant	not significant	0.0786	not significant	0.01	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant
Pyruvate	K09022	0.0829	0.0075	not significant	0.0207	not significant	0.0409	0.0896	not significant	not significant	not significant	0.054	not significant	0.0106	not significant	0.0244	not significant	0.0184	not significant	0.0129	not significant	0.0097	not significant
Pyruvate	K14155	not significant	0.0011	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0011	not significant	0.0056	0.0561	0.0227	0.0168	0.0038	not significant	0.0016	not significant
Pyruvate	K05878	not significant	0.0034	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0014	not significant	not significant	0.01	0.0177	0.0853	0.0007	not significant	0.0045	not significant
Pyruvate	K01958	not significant	0.0048	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0071	not significant	0.0044	not significant	not significant	0.0015	0.0078	not significant	0.0004	not significant	0.0143	not significant
Butyryl-CoA	K00249	not significant	0.0077	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0004	not significant	0.0897	0.04	0.0068	not significant	0.0031	not significant	0.0341	0.0886
Pyruvate	K00172	not significant	0.0084	not significant	not significant	not significant	0.0562	not significant	not significant	not significant	not significant	not significant	not significant	0.018	not significant	not significant	0.0009	0.0298	not significant	0.0654	0.021	0.0008	not significant
Acetate	K01646	not significant	0.0097	not significant	not significant	not significant	not significant	not significant	not significant	0.0601	not significant	0.0504	not significant	not significant	0.0144	0.0277	not significant	0.0343	not significant	0.0292	not significant	0.0186	not significant
Acetate	K01034	not significant	0.0102	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0029	not significant	0.0674	0.0702	0.0404	not significant	0.0156	not significant	0.0863	0.0536
Butyryl-CoA	K00209	not significant	0.0108	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0128	not significant	not significant	0.0083	0.0194	not significant	0.0142	not significant	0.0735	0.0692
Acetyl-CoA	K00059	not significant	0.0131	not significant	not significant	0.0992	not significant	0.0993	not significant	not significant	not significant	not significant	not significant	0.0583	not significant	0.0157	not significant	0.0065	not significant	0.0254	not significant	not significant	0.0608
Pyruvate	K00170	not significant	0.0153	not significant	not significant	not significant	0.0493	not significant	not significant	not significant	not significant	not significant	not significant	0.0469	not significant	not significant	0.0038	0.0287	not significant	0.0929	0.0311	0.0017	not significant
Pyruvate	K01714	not significant	0.0157	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0883	0.0866	0.022	not significant	0.0279	not significant	0.0435	not significant	0.0454	not significant
Pyruvate	K00382	not significant	0.0167	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0339	not significant	not significant	0.0409	0.013	not significant	0.0164	not significant	0.0401	not significant
Acetyl-CoA	K00640	not significant	0.0205	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0306	not significant	0.0222	not significant	0.0128	not significant	0.0178	not significant	not significant	0.0458
Acetate	K01443	not significant	0.0221	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0422	not significant	0.0924	not significant	0.0214	not significant	0.0432	not significant	not significant	0.0369
Acetyl-CoA	K04072	not significant	0.0302	not significant	not significant	not significant	not significant	0.0968	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0179	not significant	0.0459	not significant	0.0686	not significant	not significant	0.0068
Pyruvate	K00171	not significant	0.0307	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0436	not significant	not significant	0.0043	0.053	not significant	not significant	0.0446	0.0019	not significant
Acetyl-CoA	K00140	not significant	0.0309	not significant	not significant	0.0803	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0485	not significant	0.012	not significant	0.076	not significant	0.0912	not significant
Acetate	K01739	not significant	0.0338	not significant	0.0054	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0665	not significant	not significant	not significant	0.0925	not significant	not significant	0.0904
Acetate	K01060	not significant	0.0528	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0222	not significant	not significant	0.0612	not significant	not significant	not significant	not significant	0.0057	not significant
Acetate	K01644	not significant	0.053	not significant	not significant	0.0654	not significant	not significant	not significant	0.0925	not significant	not significant	not significant	not significant	not significant	0.0308	not significant	0.0683	not significant	not significant	not significant	0.061	not significant
Acetate	K00925	not significant	0.0573	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.044	not significant	0.0168	not significant	0.0973	not significant	0.0775	not significant	not significant	0.0428
Pyruvate	K05879	not significant	0.0609	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.02	0.0793	not significant	0.0116	not significant	0.0987	not significant
Pyruvate	K03737	not significant	0.0664	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0698	not significant	0.0808	not significant	not significant	not significant	not significant	0.0705
Acetate	K01740	not significant	0.0939	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0538	not significant	0.0701	not significant	0.0925	not significant	not significant	not significant	not significant	0.0669

Code

# prepare the base dataset for the plot
dat_SCFAsignif <- dat_SCFAresults %>% 
  filter((freshInt_pvalue_correct < 0.1) | (pastInt_pvalue_correct < 0.1))
dat_plot <- data.frame(SCFA         = rep(dat_SCFAsignif$SCFA,  times = 2),
                       gene         = rep(dat_SCFAsignif$gene,  times = 2),
                       baseline_medianTPMAbd = rep(dat_SCFAsignif$baseline_medianTPMAbd, times = 2),
                       baseline_prevalence    = rep(dat_SCFAsignif$baseline_prevalence,    times = 2),
                       postFresh_medianTPMAbd = rep(dat_SCFAsignif$postFresh_medianTPMAbd, times = 2),
                       postFresh_prevalence   = rep(dat_SCFAsignif$postFresh_prevalence,   times = 2),
                       postPast_medianTPMAbd  = rep(dat_SCFAsignif$postPast_medianTPMAbd,  times = 2),
                       postPast_prevalence    = rep(dat_SCFAsignif$postPast_prevalence,    times = 2),
                       model        = rep(dat_SCFAsignif$model, times = 2),
                       intervention = rep(c("Fresh intervention", "Pasteurized intervention"), each = nrow(dat_SCFAsignif)),
                       effect       = c(dat_SCFAsignif$freshInt_effect, dat_SCFAsignif$pastInt_effect),
                       qvalue       = c(dat_SCFAsignif$freshInt_pvalue_correct, dat_SCFAsignif$pastInt_pvalue_correct)) %>% 
  arrange(SCFA, desc(effect)) %>% 
  mutate(pvalue_signif = case_when((qvalue < 0.1) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (qvalue < 0.1) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (qvalue < 0.1) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (qvalue < 0.1) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model == "Gaussian") & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                   (model == "Gaussian") & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                   (model == "Gaussian") & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                   (model == "Gaussian") & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                   (model == "Gaussian") & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                   (model == "Gaussian") & (effect >= 0)        ~ "+ <0.0001 TPM",
                                   (model == "Gaussian") & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                   (model == "Gaussian") & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                   (model == "Gaussian") & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                   (model == "Gaussian") & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                   (model == "Gaussian") & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                   (model == "Gaussian") & (effect < 0)                   ~ "- >-0.0001 TPM",
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model == "log-Gaussian") & (effect >= 0)                                ~ "+ <0.01%",
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model == "log-Gaussian") & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$gene)), function(gen) {
  dat_gene <- dat_plot %>% filter(gene == gen)
  dat_gene %>% 
    dplyr::bind_rows(data.frame(SCFA          = dat_gene$SCFA[1],
                                gene          = gen,
                                intervention  = "Baseline median TPM abd.\n(prevalence)",
                                pvalue_signif = "",
                                effect_label_TPMAbd = case_when(dat_gene$baseline_medianTPMAbd[1] < .0001 ~ "<0.0001 TPM",
                                                                dat_gene$baseline_medianTPMAbd[1] < .01   ~ paste0(round(dat_gene$baseline_medianTPMAbd[1], 4), " TPM"),
                                                                dat_gene$baseline_medianTPMAbd[1] < .1    ~ paste0(round(dat_gene$baseline_medianTPMAbd[1], 3), " TPM"),
                                                                dat_gene$baseline_medianTPMAbd[1] < 1     ~ paste0(round(dat_gene$baseline_medianTPMAbd[1], 2), " TPM"),
                                                                dat_gene$baseline_medianTPMAbd[1] < 10    ~ paste0(round(dat_gene$baseline_medianTPMAbd[1], 1), " TPM"),
                                                                TRUE ~ paste0(round(dat_gene$baseline_medianTPMAbd[1], 0), " TPM")),
                                effect_label_prevalence = case_when(dat_gene$baseline_prevalence[1] < 0.01 ~ paste0(round(100 * dat_gene$baseline_prevalence[1], 1), "%"),
                                                                    TRUE                                   ~ paste0(round(100 * dat_gene$baseline_prevalence[1], 0), "%"))) %>% 
                       mutate(effect_label = paste0(effect_label_TPMAbd, "\n(", effect_label_prevalence, ")"))) %>% 
    dplyr::bind_rows(data.frame(SCFA          = dat_gene$SCFA[1],
                                gene          = gen,
                                intervention  = "After fresh median TPM abd.\n(prevalence)",
                                pvalue_signif = "",
                                effect_label_TPMAbd = case_when(dat_gene$postFresh_medianTPMAbd[1] < .0001 ~ "<0.0001 TPM",
                                                                dat_gene$postFresh_medianTPMAbd[1] < .01   ~ paste0(round(dat_gene$postFresh_medianTPMAbd[1], 4), " TPM"),
                                                                dat_gene$postFresh_medianTPMAbd[1] < .1    ~ paste0(round(dat_gene$postFresh_medianTPMAbd[1], 3), " TPM"),
                                                                dat_gene$postFresh_medianTPMAbd[1] < 1     ~ paste0(round(dat_gene$postFresh_medianTPMAbd[1], 2), " TPM"),
                                                                dat_gene$postFresh_medianTPMAbd[1] < 10    ~ paste0(round(dat_gene$postFresh_medianTPMAbd[1], 1), " TPM"),
                                                                TRUE ~ paste0(round(dat_gene$postFresh_medianTPMAbd[1], 0), " TPM")),
                                effect_label_prevalence = case_when(dat_gene$postFresh_prevalence[1] < 0.01 ~ paste0(round(100 * dat_gene$postFresh_prevalence[1], 1), "%"),
                                                                    TRUE                                    ~ paste0(round(100 * dat_gene$postFresh_prevalence[1], 0), "%"))) %>%
                       mutate(effect_label = paste0(effect_label_TPMAbd, "\n(", effect_label_prevalence, ")"))) %>%
    dplyr::bind_rows(data.frame(SCFA          = dat_gene$SCFA[1],
                                gene          = gen,
                                intervention  = "After past. median TPM abd.\n(prevalence)",
                                pvalue_signif = "",
                                effect_label_TPMAbd = case_when(dat_gene$postPast_medianTPMAbd[1] < .0001 ~ "<0.0001 TPM",
                                                                dat_gene$postPast_medianTPMAbd[1] < .01   ~ paste0(round(dat_gene$postPast_medianTPMAbd[1], 4), " TPM"),
                                                                dat_gene$postPast_medianTPMAbd[1] < .1    ~ paste0(round(dat_gene$postPast_medianTPMAbd[1], 3), " TPM"),
                                                                dat_gene$postPast_medianTPMAbd[1] < 1     ~ paste0(round(dat_gene$postPast_medianTPMAbd[1], 2), " TPM"),
                                                                dat_gene$postPast_medianTPMAbd[1] < 10    ~ paste0(round(dat_gene$postPast_medianTPMAbd[1], 1), " TPM"),
                                                                TRUE ~ paste0(round(dat_gene$postPast_medianTPMAbd[1], 0), " TPM")),
                                effect_label_prevalence = case_when(dat_gene$postPast_prevalence[1] < 0.01 ~ paste0(round(100 * dat_gene$postPast_prevalence[1], 1), "%"),
                                                                    TRUE                                   ~ paste0(round(100 * dat_gene$postPast_prevalence[1], 0), "%"))) %>%
                       mutate(effect_label = paste0(effect_label_TPMAbd, "\n(", effect_label_prevalence, ")")))
})
dat_plot_overall <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(gene         = factor(gene, levels = rev(unique(gene))),
         intervention = factor(intervention, levels = c("Baseline median TPM abd.\n(prevalence)",
                                                        "After fresh median TPM abd.\n(prevalence)",
                                                        "After past. median TPM abd.\n(prevalence)",
                                                        "Fresh intervention", "Pasteurized intervention")),
         text_size    = case_when(grepl("intervention", intervention) ~ 4,
                                  TRUE                                ~ 3),
         pvalue_signif = factor(pvalue_signif, levels = c("not significant",
                                                          "significant pos. effect (fresh intervention)",
                                                          "significant neg. effect (fresh intervention)",
                                                          "significant pos. effect (pasteurized intervention)",
                                                          "significant neg. effect (pasteurized intervention)",
                                                          "")))

# plot
dat_plot_overall %>% 
  ggplot(aes(x = intervention, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("q-value", values = c("not significant" = "gray95",
                                          "significant pos. effect (fresh intervention)" = "#7cbde9",
                                          "significant neg. effect (fresh intervention)" = "#1F78B4",
                                          "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                          "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(SCFA), scales = "free", space = "free") +
  ggtitle("Intervention effects on SCFA-related genes") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"))

Code

# ggsave("functional_geneScreening_qvalues.png", width = 10, height = 3)

Code

# prepare the base dataset for the plot
dat_SCFAsignif <- dat_SCFAresults %>% 
  filter((freshInt_pvalue < 0.05) | (pastInt_pvalue < 0.05))
dat_plot <- data.frame(SCFA         = rep(dat_SCFAsignif$SCFA,  times = 2),
                       gene         = rep(dat_SCFAsignif$gene,  times = 2),
                       baseline_medianTPMAbd = rep(dat_SCFAsignif$baseline_medianTPMAbd, times = 2),
                       model        = rep(dat_SCFAsignif$model, times = 2),
                       intervention = rep(c("Fresh intervention", "Pasteurized intervention"), each = nrow(dat_SCFAsignif)),
                       effect       = c(dat_SCFAsignif$freshInt_effect, dat_SCFAsignif$pastInt_effect),
                       pvalue       = c(dat_SCFAsignif$freshInt_pvalue, dat_SCFAsignif$pastInt_pvalue)) %>% 
  arrange(SCFA, desc(effect)) %>% 
  mutate(pvalue_signif = case_when((pvalue < 0.05) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (pvalue < 0.05) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (pvalue < 0.05) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (pvalue < 0.05) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model == "Gaussian") & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                   (model == "Gaussian") & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                   (model == "Gaussian") & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                   (model == "Gaussian") & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                   (model == "Gaussian") & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                   (model == "Gaussian") & (effect >= 0)        ~ "+ <0.0001 TPM",
                                   (model == "Gaussian") & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                   (model == "Gaussian") & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                   (model == "Gaussian") & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                   (model == "Gaussian") & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                   (model == "Gaussian") & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                   (model == "Gaussian") & (effect < 0)                   ~ "- >-0.0001 TPM",
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model == "log-Gaussian") & (effect >= 0)                                ~ "+ <0.01%",
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model == "log-Gaussian") & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$gene)), function(gen) {
  dat_gene <- dat_plot %>% filter(gene == gen)
  dat_gene %>% 
    dplyr::bind_rows(data.frame(SCFA          = dat_gene$SCFA[1],
                                gene          = gen,
                                intervention  = "Baseline median TPM abd.",
                                pvalue_signif = "",
                                effect_label  = case_when(dat_gene$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                          dat_gene$baseline_medianTPMAbd < .001  ~ paste0(round(dat_gene$baseline_medianTPMAbd, 4), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .01   ~ paste0(round(dat_gene$baseline_medianTPMAbd, 3), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .1    ~ paste0(round(dat_gene$baseline_medianTPMAbd, 2), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < 1     ~ paste0(round(dat_gene$baseline_medianTPMAbd, 1), " TPM"),
                                                          TRUE ~ paste0(round(dat_gene$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(gene          = factor(gene, levels = rev(unique(gene))),
         intervention  = factor(intervention, levels = c("Baseline median TPM abd.", "Fresh intervention", "Pasteurized intervention")),
         pvalue_signif = factor(pvalue_signif, levels = c("not significant",
                                                          "significant pos. effect (fresh intervention)",
                                                          "significant neg. effect (fresh intervention)",
                                                          "significant pos. effect (pasteurized intervention)",
                                                          "significant neg. effect (pasteurized intervention)",
                                                          "")))

# plot
dat_plot %>% 
  ggplot(aes(x = intervention, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("uncorrected p-value", values = c("not significant" = "gray95",
                                                      "significant pos. effect (fresh intervention)" = "#7cbde9",
                                                      "significant neg. effect (fresh intervention)" = "#1F78B4",
                                                      "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                                      "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(SCFA), scales = "free", space = "free") +
  ggtitle("Intervention effects on SCFA-related genes") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"))

Results for interaction effects

Overview of the numbers of significant genes

Code

data.frame("what"  = c("No. of SCFA genes",
                       "... that didn't have low abundance",
                       "No. of significant genes (fresh intervention - age low)",
                       "No. of significant genes (fresh intervention - age high)",
                       "No. of significant genes (past. intervention - age low)",
                       "No. of significant genes (past. intervention - age high)",
                       "No. of significant genes (fresh intervention - gender male)",
                       "No. of significant genes (fresh intervention - gender female)",
                       "No. of significant genes (past. intervention - gender male)",
                       "No. of significant genes (past. intervention - gender female)",
                       "No. of significant genes (fresh intervention - BMI low)",
                       "No. of significant genes (fresh intervention - BMI high)",
                       "No. of significant genes (past. intervention - BMI low)",
                       "No. of significant genes (past. intervention - BMI high)",
                       "No. of significant genes (fresh intervention - fiber low)",
                       "No. of significant genes (fresh intervention - fiber high)",
                       "No. of significant genes (past. intervention - fiber low)",
                       "No. of significant genes (past. intervention - fiber high)",
                       "No. of significant genes (fresh intervention - baseline div. low)",
                       "No. of significant genes (fresh intervention - baseline div. high)",
                       "No. of significant genes (past. intervention - baseline div. low)",
                       "No. of significant genes (past. intervention - baseline div. high)"),
           "value" = c(nrow(dat_scfaLookup),
                       nrow(dat_SCFAresults),
                       sum(dat_SCFAresults$freshInt_ageLow_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_ageHigh_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_ageLow_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_ageHigh_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_sexM_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_sexW_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_sexM_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_sexW_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_bmiLow_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_bmiHigh_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_bmiLow_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_bmiHigh_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_fibLow_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_fibHigh_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_fibLow_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_fibHigh_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_divLow_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_divHigh_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_divLow_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_divHigh_pvalue_correct < 0.1, na.rm = TRUE))) %>% 
  kable() %>% 
  kable_styling()

what	value
No. of SCFA genes	193
... that didn't have low abundance	89
No. of significant genes (fresh intervention - age low)	0
No. of significant genes (fresh intervention - age high)	0
No. of significant genes (past. intervention - age low)	2
No. of significant genes (past. intervention - age high)	0
No. of significant genes (fresh intervention - gender male)	0
No. of significant genes (fresh intervention - gender female)	0
No. of significant genes (past. intervention - gender male)	0
No. of significant genes (past. intervention - gender female)	1
No. of significant genes (fresh intervention - BMI low)	0
No. of significant genes (fresh intervention - BMI high)	0
No. of significant genes (past. intervention - BMI low)	1
No. of significant genes (past. intervention - BMI high)	0
No. of significant genes (fresh intervention - fiber low)	0
No. of significant genes (fresh intervention - fiber high)	0
No. of significant genes (past. intervention - fiber low)	3
No. of significant genes (past. intervention - fiber high)	0
No. of significant genes (fresh intervention - baseline div. low)	0
No. of significant genes (fresh intervention - baseline div. high)	0
No. of significant genes (past. intervention - baseline div. low)	0
No. of significant genes (past. intervention - baseline div. high)	0

… alternatively looking at the uncorrected p-values

Code

data.frame("what"  = c("No. of SCFA genes",
                       "... that didn't have low abundance",
                       "No. of significant genes (fresh intervention - age low)",
                       "No. of significant genes (fresh intervention - age high)",
                       "No. of significant genes (past. intervention - age low)",
                       "No. of significant genes (past. intervention - age high)",
                       "No. of significant genes (fresh intervention - gender male)",
                       "No. of significant genes (fresh intervention - gender female)",
                       "No. of significant genes (past. intervention - gender male)",
                       "No. of significant genes (past. intervention - gender female)",
                       "No. of significant genes (fresh intervention - BMI low)",
                       "No. of significant genes (fresh intervention - BMI high)",
                       "No. of significant genes (past. intervention - BMI low)",
                       "No. of significant genes (past. intervention - BMI high)",
                       "No. of significant genes (fresh intervention - fiber low)",
                       "No. of significant genes (fresh intervention - fiber high)",
                       "No. of significant genes (past. intervention - fiber low)",
                       "No. of significant genes (past. intervention - fiber high)",
                       "No. of significant genes (fresh intervention - baseline div. low)",
                       "No. of significant genes (fresh intervention - baseline div. high)",
                       "No. of significant genes (past. intervention - baseline div. low)",
                       "No. of significant genes (past. intervention - baseline div. high)"),
           "value" = c(nrow(dat_scfaLookup),
                       nrow(dat_SCFAresults),
                       sum(dat_SCFAresults$freshInt_ageLow_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_ageHigh_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_ageLow_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_ageHigh_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_sexM_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_sexW_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_sexM_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_sexW_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_bmiLow_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_bmiHigh_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_bmiLow_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_bmiHigh_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_fibLow_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_fibHigh_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_fibLow_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_fibHigh_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_divLow_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$freshInt_divHigh_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_divLow_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_SCFAresults$pastInt_divHigh_pvalue < 0.05, na.rm = TRUE))) %>% 
  kable() %>% 
  kable_styling()

what	value
No. of SCFA genes	193
... that didn't have low abundance	89
No. of significant genes (fresh intervention - age low)	1
No. of significant genes (fresh intervention - age high)	5
No. of significant genes (past. intervention - age low)	21
No. of significant genes (past. intervention - age high)	1
No. of significant genes (fresh intervention - gender male)	1
No. of significant genes (fresh intervention - gender female)	4
No. of significant genes (past. intervention - gender male)	12
No. of significant genes (past. intervention - gender female)	11
No. of significant genes (fresh intervention - BMI low)	2
No. of significant genes (fresh intervention - BMI high)	2
No. of significant genes (past. intervention - BMI low)	23
No. of significant genes (past. intervention - BMI high)	1
No. of significant genes (fresh intervention - fiber low)	1
No. of significant genes (fresh intervention - fiber high)	7
No. of significant genes (past. intervention - fiber low)	15
No. of significant genes (past. intervention - fiber high)	4
No. of significant genes (fresh intervention - baseline div. low)	10
No. of significant genes (fresh intervention - baseline div. high)	0
No. of significant genes (past. intervention - baseline div. low)	15
No. of significant genes (past. intervention - baseline div. high)	6

baseline diversity

Code

# prepare the base dataset for the plot
dat_signif <- dat_SCFAresults %>% 
  filter((freshInt_divLow_pvalue_correct < 0.1) | (freshInt_divHigh_pvalue_correct < 0.1) |
           (pastInt_divLow_pvalue_correct < 0.1) | (pastInt_divHigh_pvalue_correct < 0.1))
if (nrow(dat_signif) == 0) {
  message("No corrected p-value < 0.1")
} else {
  dat_plot <- data.frame(SCFA          = rep(dat_signif$SCFA,  times = 2),
                         gene          = rep(dat_signif$gene,  times = 2),
                         baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                         model         = rep(dat_signif$model, times = 4),
                         intervention  = rep(c("Fresh intervention\n(baseline div. low)", "Fresh intervention\n(baseline div. high)",
                                               "Pasteurized intervention\n(baseline div. low)", "Pasteurized intervention\n(baseline div. high)"), each = nrow(dat_signif)),
                         effect        = c(dat_signif$freshInt_divLow_effect, dat_signif$freshInt_divHigh_effect,
                                           dat_signif$pastInt_divLow_effect,  dat_signif$pastInt_divHigh_effect),
                         qvalue        = c(dat_signif$freshInt_divLow_pvalue_correct, dat_signif$freshInt_divHigh_pvalue_correct,
                                           dat_signif$pastInt_divLow_pvalue_correct,  dat_signif$pastInt_divHigh_pvalue_correct),
                         pastEffect_forSorting = rep(dat_signif$pastInt_divLow_effect, times = 4)) %>% 
    arrange(SCFA, desc(pastEffect_forSorting)) %>% 
    mutate(pvalue_signif = case_when((qvalue < 0.1) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                     (qvalue < 0.1) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                     (qvalue < 0.1) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                     (qvalue < 0.1) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                     TRUE          ~ "not significant"),
           effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                     (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                     (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                     (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                     (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                     (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 TPM",
                                     (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                     (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                     (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                     (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                     (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                     (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 TPM",
                                     (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                     (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                     (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                     (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                     (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                     (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                     (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                     (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))
  
  # artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
  dat_plot_list <- lapply(as.character(unique(dat_plot$gene)), function(gen) {
    dat_gene <- dat_plot %>% filter(gene == gen)
    dat_gene %>% 
      dplyr::bind_rows(data.frame(SCFA          = dat_gene$SCFA[1],
                                  gene          = gen,
                                  intervention  = "Baseline median TPM abd.",
                                  pvalue_signif = "",
                                  effect_label  = case_when(dat_gene$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                            dat_gene$baseline_medianTPMAbd < .001  ~ paste0(round(dat_gene$baseline_medianTPMAbd, 4), " TPM"),
                                                            dat_gene$baseline_medianTPMAbd < .01   ~ paste0(round(dat_gene$baseline_medianTPMAbd, 3), " TPM"),
                                                            dat_gene$baseline_medianTPMAbd < .1    ~ paste0(round(dat_gene$baseline_medianTPMAbd, 2), " TPM"),
                                                            dat_gene$baseline_medianTPMAbd < 1     ~ paste0(round(dat_gene$baseline_medianTPMAbd, 1), " TPM"),
                                                            TRUE ~ paste0(round(dat_gene$baseline_medianTPMAbd, 0), " TPM"))))
  })
  dat_plot_divInt <- dat_plot_list %>% dplyr::bind_rows() %>% 
    mutate(gene         = factor(gene, levels = rev(unique(gene))),
           intervention = factor(intervention, levels = c("Baseline median TPM abd.", "Fresh intervention\n(baseline div. low)", "Fresh intervention\n(baseline div. high)",
                                                          "Pasteurized intervention\n(baseline div. low)", "Pasteurized intervention\n(baseline div. high)")),
           pvalue_signif = factor(pvalue_signif, levels = c("not significant",
                                                            "significant pos. effect (fresh intervention)",
                                                            "significant neg. effect (fresh intervention)",
                                                            "significant pos. effect (pasteurized intervention)",
                                                            "significant neg. effect (pasteurized intervention)",
                                                            "")))
  
  # plot
  dat_plot_divInt %>% 
    ggplot(aes(x = intervention, y = gene)) +
    geom_tile(aes(fill = pvalue_signif)) +
    geom_text(aes(label = effect_label)) +
    scale_fill_manual("q-value", values = c("not significant" = "gray95",
                                            "significant pos. effect (fresh intervention)" = "#7cbde9",
                                            "significant neg. effect (fresh intervention)" = "#1F78B4",
                                            "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                            "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                      na.value = "white") +
    facet_grid(rows = vars(SCFA), scales = "free", space = "free") +
    ggtitle("Intervention effects on SCFA genes") +
    theme(axis.title = element_blank(),
          panel.grid = element_blank(),
          strip.background = element_rect(fill = "gray97", color = "gray97"))
  # ggsave("functional_geneScreening_divInteractions_qvalues.png", width = 12, height = 1.5)
}

No corrected p-value < 0.1

Code

# prepare the base dataset for the plot
dat_signif <- dat_SCFAresults %>% 
  filter((freshInt_divLow_pvalue < 0.05) | (freshInt_divHigh_pvalue < 0.05) |
           (pastInt_divLow_pvalue < 0.05) | (pastInt_divHigh_pvalue < 0.05))
dat_plot <- data.frame(SCFA          = rep(dat_signif$SCFA,  times = 2),
                       gene          = rep(dat_signif$gene,  times = 2),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(baseline div. low)", "Fresh intervention\n(baseline div. high)",
                                             "Pasteurized intervention\n(baseline div. low)", "Pasteurized intervention\n(baseline div. high)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_divLow_effect, dat_signif$freshInt_divHigh_effect,
                                         dat_signif$pastInt_divLow_effect,  dat_signif$pastInt_divHigh_effect),
                       pvalue        = c(dat_signif$freshInt_divLow_pvalue, dat_signif$freshInt_divHigh_pvalue,
                                         dat_signif$pastInt_divLow_pvalue,  dat_signif$pastInt_divHigh_pvalue),
                       pastEffect_forSorting = rep(dat_signif$pastInt_divLow_effect, times = 4)) %>% 
  arrange(SCFA, desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((pvalue < 0.05) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (pvalue < 0.05) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (pvalue < 0.05) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (pvalue < 0.05) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 TPM",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 TPM",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$gene)), function(gen) {
  dat_gene <- dat_plot %>% filter(gene == gen)
  dat_gene %>% 
    dplyr::bind_rows(data.frame(SCFA          = dat_gene$SCFA[1],
                                gene          = gen,
                                intervention  = "Baseline median TPM abd.",
                                pvalue_signif = "",
                                effect_label  = case_when(dat_gene$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                          dat_gene$baseline_medianTPMAbd < .001  ~ paste0(round(dat_gene$baseline_medianTPMAbd, 4), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .01   ~ paste0(round(dat_gene$baseline_medianTPMAbd, 3), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .1    ~ paste0(round(dat_gene$baseline_medianTPMAbd, 2), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < 1     ~ paste0(round(dat_gene$baseline_medianTPMAbd, 1), " TPM"),
                                                          TRUE ~ paste0(round(dat_gene$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(gene          = factor(gene, levels = rev(unique(gene))),
         intervention  = gsub(intervention, pattern = "low",  replacement = "[2.8, 3.85)"),
         intervention  = gsub(intervention, pattern = "high", replacement = "[3.85, 4.4)"),
         intervention  = factor(intervention, levels = c("Baseline median TPM abd.",
                                                         "Fresh intervention\n(baseline div. [2.8, 3.85))",
                                                         "Fresh intervention\n(baseline div. [3.85, 4.4))",
                                                         "Pasteurized intervention\n(baseline div. [2.8, 3.85))",
                                                         "Pasteurized intervention\n(baseline div. [3.85, 4.4))")),
         pvalue_signif = factor(pvalue_signif, levels = c("not significant",
                                                          "significant pos. effect (fresh intervention)",
                                                          "significant neg. effect (fresh intervention)",
                                                          "significant pos. effect (pasteurized intervention)",
                                                          "significant neg. effect (pasteurized intervention)",
                                                          "")),)

# plot
dat_plot %>% 
  ggplot(aes(x = intervention, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("uncorrected p-value", values = c("not significant" = "gray95",
                                                      "significant pos. effect (fresh intervention)" = "#7cbde9",
                                                      "significant neg. effect (fresh intervention)" = "#1F78B4",
                                                      "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                                      "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(SCFA), scales = "free", space = "free") +
  ggtitle("Intervention effects on SCFA genes") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"))

age

Code

# prepare the base dataset for the plot
dat_signif <- dat_SCFAresults %>% 
  filter((freshInt_ageLow_pvalue_correct < 0.1) | (freshInt_ageHigh_pvalue_correct < 0.1) |
           (pastInt_ageLow_pvalue_correct < 0.1) | (pastInt_ageHigh_pvalue_correct < 0.1))
dat_plot <- data.frame(SCFA          = rep(dat_signif$SCFA,  times = 4),
                       gene          = rep(dat_signif$gene,  times = 4),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(age low)", "Fresh intervention\n(age high)",
                                             "Pasteurized intervention\n(age low)", "Pasteurized intervention\n(age high)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_ageLow_effect, dat_signif$freshInt_ageHigh_effect,
                                         dat_signif$pastInt_ageLow_effect,  dat_signif$pastInt_ageHigh_effect),
                       qvalue        = c(dat_signif$freshInt_ageLow_pvalue_correct, dat_signif$freshInt_ageHigh_pvalue_correct,
                                         dat_signif$pastInt_ageLow_pvalue_correct,  dat_signif$pastInt_ageHigh_pvalue_correct),
                       pastEffect_forSorting = rep(dat_signif$pastInt_ageLow_effect, times = 4)) %>% 
  arrange(SCFA, desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((qvalue < 0.1) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (qvalue < 0.1) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (qvalue < 0.1) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (qvalue < 0.1) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 TPM",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 TPM",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$gene)), function(gen) {
  dat_gene <- dat_plot %>% filter(gene == gen)
  dat_gene %>% 
    dplyr::bind_rows(data.frame(SCFA          = dat_gene$SCFA[1],
                                gene          = gen,
                                intervention  = "Baseline median TPM abd.",
                                pvalue_signif = "",
                                effect_label  = case_when(dat_gene$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                          dat_gene$baseline_medianTPMAbd < .001  ~ paste0(round(dat_gene$baseline_medianTPMAbd, 4), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .01   ~ paste0(round(dat_gene$baseline_medianTPMAbd, 3), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .1    ~ paste0(round(dat_gene$baseline_medianTPMAbd, 2), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < 1     ~ paste0(round(dat_gene$baseline_medianTPMAbd, 1), " TPM"),
                                                          TRUE ~ paste0(round(dat_gene$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot_ageInt <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(gene         = factor(gene, levels = rev(unique(gene))),
         intervention = factor(intervention, levels = c("Baseline median TPM abd.", "Fresh intervention\n(age low)", "Fresh intervention\n(age high)",
                                                        "Pasteurized intervention\n(age low)", "Pasteurized intervention\n(age high)")),
         pvalue_signif = factor(pvalue_signif, levels = c("not significant",
                                                          "significant pos. effect (fresh intervention)",
                                                          "significant neg. effect (fresh intervention)",
                                                          "significant pos. effect (pasteurized intervention)",
                                                          "significant neg. effect (pasteurized intervention)",
                                                          "")))

# plot
dat_plot_ageInt %>% 
  ggplot(aes(x = intervention, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("q-value", values = c("not significant" = "gray95",
                                          "significant pos. effect (fresh intervention)" = "#7cbde9",
                                          "significant neg. effect (fresh intervention)" = "#1F78B4",
                                          "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                          "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(SCFA), scales = "free", space = "free") +
  ggtitle("Intervention effects on SCFA genes") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"))

Code

# ggsave("functional_geneScreening_ageInteractions_qvalues.png", width = 12, height = 2)

Code

# prepare the base dataset for the plot
dat_signif <- dat_SCFAresults %>% 
  filter((freshInt_ageLow_pvalue < 0.05) | (freshInt_ageHigh_pvalue < 0.05) |
           (pastInt_ageLow_pvalue < 0.05) | (pastInt_ageHigh_pvalue < 0.05))
dat_plot <- data.frame(SCFA          = rep(dat_signif$SCFA,  times = 4),
                       gene          = rep(dat_signif$gene,  times = 4),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(age low)", "Fresh intervention\n(age high)",
                                             "Pasteurized intervention\n(age low)", "Pasteurized intervention\n(age high)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_ageLow_effect, dat_signif$freshInt_ageHigh_effect,
                                         dat_signif$pastInt_ageLow_effect,  dat_signif$pastInt_ageHigh_effect),
                       pvalue        = c(dat_signif$freshInt_ageLow_pvalue, dat_signif$freshInt_ageHigh_pvalue,
                                         dat_signif$pastInt_ageLow_pvalue,  dat_signif$pastInt_ageHigh_pvalue),
                       pastEffect_forSorting = rep(dat_signif$pastInt_ageLow_effect, times = 4)) %>% 
  arrange(SCFA, desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((pvalue < 0.05) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (pvalue < 0.05) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (pvalue < 0.05) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (pvalue < 0.05) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 TPM",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 TPM",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$gene)), function(gen) {
  dat_gene <- dat_plot %>% filter(gene == gen)
  dat_gene %>% 
    dplyr::bind_rows(data.frame(SCFA          = dat_gene$SCFA[1],
                                gene          = gen,
                                intervention  = "Baseline median TPM abd.",
                                pvalue_signif = "",
                                effect_label  = case_when(dat_gene$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                          dat_gene$baseline_medianTPMAbd < .001  ~ paste0(round(dat_gene$baseline_medianTPMAbd, 4), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .01   ~ paste0(round(dat_gene$baseline_medianTPMAbd, 3), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .1    ~ paste0(round(dat_gene$baseline_medianTPMAbd, 2), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < 1     ~ paste0(round(dat_gene$baseline_medianTPMAbd, 1), " TPM"),
                                                          TRUE ~ paste0(round(dat_gene$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(gene          = factor(gene, levels = rev(unique(gene))),
         intervention  = gsub(intervention, pattern = "age low",  replacement = "age 21-49"),
         intervention  = gsub(intervention, pattern = "age high", replacement = "age 50-69"),
         intervention  = factor(intervention, levels = c("Baseline median TPM abd.",
                                                         "Fresh intervention\n(age 21-49)",
                                                         "Fresh intervention\n(age 50-69)",
                                                         "Pasteurized intervention\n(age 21-49)",
                                                         "Pasteurized intervention\n(age 50-69)")),
         pvalue_signif = factor(pvalue_signif, levels = c("not significant",
                                                          "significant pos. effect (fresh intervention)",
                                                          "significant neg. effect (fresh intervention)",
                                                          "significant pos. effect (pasteurized intervention)",
                                                          "significant neg. effect (pasteurized intervention)",
                                                          "")),
         SCFA          = case_when(SCFA == "Acetyl-CoA" ~ "ACoA",
                                   SCFA == "succinate"  ~ "Succ.",
                                   TRUE                 ~ SCFA))

# plot
dat_plot %>% 
  ggplot(aes(x = intervention, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("uncorrected p-value", values = c("not significant" = "gray95",
                                                      "significant pos. effect (fresh intervention)" = "#7cbde9",
                                                      "significant neg. effect (fresh intervention)" = "#1F78B4",
                                                      "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                                      "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(SCFA), scales = "free", space = "free") +
  ggtitle("Intervention effects on SCFA genes") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"))

sex

Code

# prepare the base dataset for the plot
dat_signif <- dat_SCFAresults %>% 
  filter((freshInt_sexM_pvalue_correct < 0.1) | (freshInt_sexW_pvalue_correct < 0.1) |
           (pastInt_sexM_pvalue_correct < 0.1) | (pastInt_sexW_pvalue_correct < 0.1))
dat_plot <- data.frame(SCFA          = rep(dat_signif$SCFA,  times = 2),
                       gene          = rep(dat_signif$gene,  times = 2),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(gender male)", "Fresh intervention\n(gender female)",
                                             "Pasteurized intervention\n(gender male)", "Pasteurized intervention\n(gender female)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_sexM_effect, dat_signif$freshInt_sexW_effect,
                                         dat_signif$pastInt_sexM_effect,  dat_signif$pastInt_sexW_effect),
                       qvalue        = c(dat_signif$freshInt_sexM_pvalue_correct, dat_signif$freshInt_sexW_pvalue_correct,
                                         dat_signif$pastInt_sexM_pvalue_correct,  dat_signif$pastInt_sexW_pvalue_correct),
                       pastEffect_forSorting = rep(dat_signif$pastInt_sexM_effect, times = 4)) %>% 
  arrange(SCFA, desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((qvalue < 0.1) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (qvalue < 0.1) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (qvalue < 0.1) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (qvalue < 0.1) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 TPM",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 TPM",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$gene)), function(gen) {
  dat_gene <- dat_plot %>% filter(gene == gen)
  dat_gene %>% 
    dplyr::bind_rows(data.frame(SCFA          = dat_gene$SCFA[1],
                                gene          = gen,
                                intervention  = "Baseline median TPM abd.",
                                pvalue_signif = "",
                                effect_label  = case_when(dat_gene$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                          dat_gene$baseline_medianTPMAbd < .001  ~ paste0(round(dat_gene$baseline_medianTPMAbd, 4), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .01   ~ paste0(round(dat_gene$baseline_medianTPMAbd, 3), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .1    ~ paste0(round(dat_gene$baseline_medianTPMAbd, 2), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < 1     ~ paste0(round(dat_gene$baseline_medianTPMAbd, 1), " TPM"),
                                                          TRUE ~ paste0(round(dat_gene$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot_sexInt <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(gene         = factor(gene, levels = rev(unique(gene))),
         intervention = factor(intervention, levels = c("Baseline median TPM abd.", "Fresh intervention\n(gender male)", "Fresh intervention\n(gender female)",
                                                        "Pasteurized intervention\n(gender male)", "Pasteurized intervention\n(gender female)")),
         pvalue_signif = factor(pvalue_signif, levels = c("not significant",
                                                          "significant pos. effect (fresh intervention)",
                                                          "significant neg. effect (fresh intervention)",
                                                          "significant pos. effect (pasteurized intervention)",
                                                          "significant neg. effect (pasteurized intervention)",
                                                          "")))

# plot
dat_plot_sexInt %>% 
  ggplot(aes(x = intervention, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("q-value", values = c("not significant" = "gray95",
                                          "significant pos. effect (fresh intervention)" = "#7cbde9",
                                          "significant neg. effect (fresh intervention)" = "#1F78B4",
                                          "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                          "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(SCFA), scales = "free", space = "free") +
  ggtitle("Intervention effects on SCFA genes") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"))

Code

# ggsave("functional_geneScreening_sexInteractions_qvalues.png", width = 12, height = 1.5)

Code

# prepare the base dataset for the plot
dat_signif <- dat_SCFAresults %>% 
  filter((freshInt_sexM_pvalue < 0.05) | (freshInt_sexW_pvalue < 0.05) |
           (pastInt_sexM_pvalue < 0.05) | (pastInt_sexW_pvalue < 0.05))
dat_plot <- data.frame(SCFA          = rep(dat_signif$SCFA,  times = 2),
                       gene          = rep(dat_signif$gene,  times = 2),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(gender male)", "Fresh intervention\n(gender female)",
                                             "Pasteurized intervention\n(gender male)", "Pasteurized intervention\n(gender female)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_sexM_effect, dat_signif$freshInt_sexW_effect,
                                         dat_signif$pastInt_sexM_effect,  dat_signif$pastInt_sexW_effect),
                       pvalue        = c(dat_signif$freshInt_sexM_pvalue, dat_signif$freshInt_sexW_pvalue,
                                         dat_signif$pastInt_sexM_pvalue,  dat_signif$pastInt_sexW_pvalue),
                       pastEffect_forSorting = rep(dat_signif$pastInt_sexM_effect, times = 4)) %>% 
  arrange(SCFA, desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((pvalue < 0.05) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (pvalue < 0.05) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (pvalue < 0.05) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (pvalue < 0.05) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 TPM",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 TPM",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$gene)), function(gen) {
  dat_gene <- dat_plot %>% filter(gene == gen)
  dat_gene %>% 
    dplyr::bind_rows(data.frame(SCFA          = dat_gene$SCFA[1],
                                gene          = gen,
                                intervention  = "Baseline median TPM abd.",
                                pvalue_signif = "",
                                effect_label  = case_when(dat_gene$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                          dat_gene$baseline_medianTPMAbd < .001  ~ paste0(round(dat_gene$baseline_medianTPMAbd, 4), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .01   ~ paste0(round(dat_gene$baseline_medianTPMAbd, 3), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .1    ~ paste0(round(dat_gene$baseline_medianTPMAbd, 2), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < 1     ~ paste0(round(dat_gene$baseline_medianTPMAbd, 1), " TPM"),
                                                          TRUE ~ paste0(round(dat_gene$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(gene          = factor(gene, levels = rev(unique(gene))),
         intervention  = gsub(intervention, pattern = "gender ", replacement = ""),
         intervention  = factor(intervention, levels = c("Baseline median TPM abd.",
                                                         "Fresh intervention\n(male)",
                                                         "Fresh intervention\n(female)",
                                                         "Pasteurized intervention\n(male)",
                                                         "Pasteurized intervention\n(female)")),
         pvalue_signif = factor(pvalue_signif, levels = c("not significant",
                                                          "significant pos. effect (fresh intervention)",
                                                          "significant neg. effect (fresh intervention)",
                                                          "significant pos. effect (pasteurized intervention)",
                                                          "significant neg. effect (pasteurized intervention)",
                                                          "")),)

# plot
dat_plot %>% 
  ggplot(aes(x = intervention, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("uncorrected p-value", values = c("not significant" = "gray95",
                                                      "significant pos. effect (fresh intervention)" = "#7cbde9",
                                                      "significant neg. effect (fresh intervention)" = "#1F78B4",
                                                      "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                                      "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(SCFA), scales = "free", space = "free") +
  ggtitle("Intervention effects on SCFA genes") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"))

BMI

Code

# prepare the base dataset for the plot
dat_signif <- dat_SCFAresults %>% 
  filter((freshInt_bmiLow_pvalue_correct < 0.1) | (freshInt_bmiHigh_pvalue_correct < 0.1) |
           (pastInt_bmiLow_pvalue_correct < 0.1) | (pastInt_bmiHigh_pvalue_correct < 0.1))
dat_plot <- data.frame(SCFA          = rep(dat_signif$SCFA,  times = 2),
                       gene          = rep(dat_signif$gene,  times = 2),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(BMI low)", "Fresh intervention\n(BMI high)",
                                             "Pasteurized intervention\n(BMI low)", "Pasteurized intervention\n(BMI high)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_bmiLow_effect, dat_signif$freshInt_bmiHigh_effect,
                                         dat_signif$pastInt_bmiLow_effect,  dat_signif$pastInt_bmiHigh_effect),
                       qvalue        = c(dat_signif$freshInt_bmiLow_pvalue_correct, dat_signif$freshInt_bmiHigh_pvalue_correct,
                                         dat_signif$pastInt_bmiLow_pvalue_correct,  dat_signif$pastInt_bmiHigh_pvalue_correct),
                       pastEffect_forSorting = rep(dat_signif$pastInt_bmiLow_effect, times = 4)) %>% 
  arrange(SCFA, desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((qvalue < 0.1) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (qvalue < 0.1) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (qvalue < 0.1) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (qvalue < 0.1) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 TPM",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 TPM",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$gene)), function(gen) {
  dat_gene <- dat_plot %>% filter(gene == gen)
  dat_gene %>% 
    dplyr::bind_rows(data.frame(SCFA          = dat_gene$SCFA[1],
                                gene          = gen,
                                intervention  = "Baseline median TPM abd.",
                                pvalue_signif = "",
                                effect_label  = case_when(dat_gene$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                          dat_gene$baseline_medianTPMAbd < .001  ~ paste0(round(dat_gene$baseline_medianTPMAbd, 4), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .01   ~ paste0(round(dat_gene$baseline_medianTPMAbd, 3), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .1    ~ paste0(round(dat_gene$baseline_medianTPMAbd, 2), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < 1     ~ paste0(round(dat_gene$baseline_medianTPMAbd, 1), " TPM"),
                                                          TRUE ~ paste0(round(dat_gene$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot_bmiInt <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(gene         = factor(gene, levels = rev(unique(gene))),
         intervention = factor(intervention, levels = c("Baseline median TPM abd.", "Fresh intervention\n(BMI low)", "Fresh intervention\n(BMI high)",
                                                        "Pasteurized intervention\n(BMI low)", "Pasteurized intervention\n(BMI high)")),
         pvalue_signif = factor(pvalue_signif, levels = c("not significant",
                                                          "significant pos. effect (fresh intervention)",
                                                          "significant neg. effect (fresh intervention)",
                                                          "significant pos. effect (pasteurized intervention)",
                                                          "significant neg. effect (pasteurized intervention)",
                                                          "")))

# plot
dat_plot_bmiInt %>% 
  ggplot(aes(x = intervention, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("q-value", values = c("not significant" = "gray95",
                                          "significant pos. effect (fresh intervention)" = "#7cbde9",
                                          "significant neg. effect (fresh intervention)" = "#1F78B4",
                                          "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                          "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(SCFA), scales = "free", space = "free") +
  ggtitle("Intervention effects on SCFA genes") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"))

Code

# ggsave("functional_geneScreening_bmiInteractions_qvalues.png", width = 12, height = 1.5)

Code

# prepare the base dataset for the plot
dat_signif <- dat_SCFAresults %>% 
  filter((freshInt_bmiLow_pvalue < 0.05) | (freshInt_bmiHigh_pvalue < 0.05) |
           (pastInt_bmiLow_pvalue < 0.05) | (pastInt_bmiHigh_pvalue < 0.05))
dat_plot <- data.frame(SCFA          = rep(dat_signif$SCFA,  times = 2),
                       gene          = rep(dat_signif$gene,  times = 2),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(BMI low)", "Fresh intervention\n(BMI high)",
                                             "Pasteurized intervention\n(BMI low)", "Pasteurized intervention\n(BMI high)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_bmiLow_effect, dat_signif$freshInt_bmiHigh_effect,
                                         dat_signif$pastInt_bmiLow_effect,  dat_signif$pastInt_bmiHigh_effect),
                       pvalue        = c(dat_signif$freshInt_bmiLow_pvalue, dat_signif$freshInt_bmiHigh_pvalue,
                                         dat_signif$pastInt_bmiLow_pvalue,  dat_signif$pastInt_bmiHigh_pvalue),
                       pastEffect_forSorting = rep(dat_signif$pastInt_bmiLow_effect, times = 4)) %>% 
  arrange(SCFA, desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((pvalue < 0.05) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (pvalue < 0.05) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (pvalue < 0.05) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (pvalue < 0.05) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 TPM",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 TPM",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$gene)), function(gen) {
  dat_gene <- dat_plot %>% filter(gene == gen)
  dat_gene %>% 
    dplyr::bind_rows(data.frame(SCFA          = dat_gene$SCFA[1],
                                gene          = gen,
                                intervention  = "Baseline median TPM abd.",
                                pvalue_signif = "",
                                effect_label  = case_when(dat_gene$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                          dat_gene$baseline_medianTPMAbd < .001  ~ paste0(round(dat_gene$baseline_medianTPMAbd, 4), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .01   ~ paste0(round(dat_gene$baseline_medianTPMAbd, 3), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .1    ~ paste0(round(dat_gene$baseline_medianTPMAbd, 2), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < 1     ~ paste0(round(dat_gene$baseline_medianTPMAbd, 1), " TPM"),
                                                          TRUE ~ paste0(round(dat_gene$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(gene          = factor(gene, levels = rev(unique(gene))),
         intervention  = gsub(intervention, pattern = "low",  replacement = "[18, 25)"),
         intervention  = gsub(intervention, pattern = "high", replacement = "[25, 31]"),
         intervention  = factor(intervention, levels = c("Baseline median TPM abd.",
                                                         "Fresh intervention\n(BMI [18, 25))",
                                                         "Fresh intervention\n(BMI [25, 31])",
                                                         "Pasteurized intervention\n(BMI [18, 25))",
                                                         "Pasteurized intervention\n(BMI [25, 31])")),
         pvalue_signif = factor(pvalue_signif, levels = c("not significant",
                                                          "significant pos. effect (fresh intervention)",
                                                          "significant neg. effect (fresh intervention)",
                                                          "significant pos. effect (pasteurized intervention)",
                                                          "significant neg. effect (pasteurized intervention)",
                                                          "")),)

# plot
dat_plot %>% 
  ggplot(aes(x = intervention, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("uncorrected p-value", values = c("not significant" = "gray95",
                                                      "significant pos. effect (fresh intervention)" = "#7cbde9",
                                                      "significant neg. effect (fresh intervention)" = "#1F78B4",
                                                      "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                                      "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(SCFA), scales = "free", space = "free") +
  ggtitle("Intervention effects on SCFA genes") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"))

fiber

Code

# prepare the base dataset for the plot
dat_signif <- dat_SCFAresults %>% 
  filter((freshInt_fibLow_pvalue_correct < 0.1) | (freshInt_fibHigh_pvalue_correct < 0.1) |
           (pastInt_fibLow_pvalue_correct < 0.1) | (pastInt_fibHigh_pvalue_correct < 0.1))
dat_plot <- data.frame(SCFA          = rep(dat_signif$SCFA,  times = 2),
                       gene          = rep(dat_signif$gene,  times = 2),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(fiber low)", "Fresh intervention\n(fiber high)",
                                             "Pasteurized intervention\n(fiber low)", "Pasteurized intervention\n(fiber high)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_fibLow_effect, dat_signif$freshInt_fibHigh_effect,
                                         dat_signif$pastInt_fibLow_effect,  dat_signif$pastInt_fibHigh_effect),
                       qvalue        = c(dat_signif$freshInt_fibLow_pvalue_correct, dat_signif$freshInt_fibHigh_pvalue_correct,
                                         dat_signif$pastInt_fibLow_pvalue_correct,  dat_signif$pastInt_fibHigh_pvalue_correct),
                       pastEffect_forSorting = rep(dat_signif$pastInt_fibLow_effect, times = 4)) %>% 
  arrange(SCFA, desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((qvalue < 0.1) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (qvalue < 0.1) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (qvalue < 0.1) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (qvalue < 0.1) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 TPM",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 TPM",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$gene)), function(gen) {
  dat_gene <- dat_plot %>% filter(gene == gen)
  dat_gene %>% 
    dplyr::bind_rows(data.frame(SCFA          = dat_gene$SCFA[1],
                                gene          = gen,
                                intervention  = "Baseline median TPM abd.",
                                pvalue_signif = "",
                                effect_label  = case_when(dat_gene$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                          dat_gene$baseline_medianTPMAbd < .001  ~ paste0(round(dat_gene$baseline_medianTPMAbd, 4), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .01   ~ paste0(round(dat_gene$baseline_medianTPMAbd, 3), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .1    ~ paste0(round(dat_gene$baseline_medianTPMAbd, 2), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < 1     ~ paste0(round(dat_gene$baseline_medianTPMAbd, 1), " TPM"),
                                                          TRUE ~ paste0(round(dat_gene$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot_fibInt <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(gene         = factor(gene, levels = rev(unique(gene))),
         intervention = factor(intervention, levels = c("Baseline median TPM abd.", "Fresh intervention\n(fiber low)", "Fresh intervention\n(fiber high)",
                                                        "Pasteurized intervention\n(fiber low)", "Pasteurized intervention\n(fiber high)")),
         pvalue_signif = factor(pvalue_signif, levels = c("not significant",
                                                          "significant pos. effect (fresh intervention)",
                                                          "significant neg. effect (fresh intervention)",
                                                          "significant pos. effect (pasteurized intervention)",
                                                          "significant neg. effect (pasteurized intervention)",
                                                          "")))

# plot
dat_plot_fibInt %>% 
  ggplot(aes(x = intervention, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("q-value", values = c("not significant" = "gray95",
                                          "significant pos. effect (fresh intervention)" = "#7cbde9",
                                          "significant neg. effect (fresh intervention)" = "#1F78B4",
                                          "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                          "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(SCFA), scales = "free", space = "free") +
  ggtitle("Intervention effects on SCFA genes") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"))

Code

# ggsave("functional_geneScreening_fibInteractions_qvalues.png", width = 12, height = 1.5)

Code

# prepare the base dataset for the plot
dat_signif <- dat_SCFAresults %>% 
  filter((freshInt_fibLow_pvalue < 0.05) | (freshInt_fibHigh_pvalue < 0.05) |
           (pastInt_fibLow_pvalue < 0.05) | (pastInt_fibHigh_pvalue < 0.05))
dat_plot <- data.frame(SCFA          = rep(dat_signif$SCFA,  times = 2),
                       gene          = rep(dat_signif$gene,  times = 2),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(fiber low)", "Fresh intervention\n(fiber high)",
                                             "Pasteurized intervention\n(fiber low)", "Pasteurized intervention\n(fiber high)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_fibLow_effect, dat_signif$freshInt_fibHigh_effect,
                                         dat_signif$pastInt_fibLow_effect,  dat_signif$pastInt_fibHigh_effect),
                       pvalue        = c(dat_signif$freshInt_fibLow_pvalue, dat_signif$freshInt_fibHigh_pvalue,
                                         dat_signif$pastInt_fibLow_pvalue,  dat_signif$pastInt_fibHigh_pvalue),
                       pastEffect_forSorting = rep(dat_signif$pastInt_fibLow_effect, times = 4)) %>% 
  arrange(SCFA, desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((pvalue < 0.05) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (pvalue < 0.05) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (pvalue < 0.05) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (pvalue < 0.05) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 TPM",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 TPM",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$gene)), function(gen) {
  dat_gene <- dat_plot %>% filter(gene == gen)
  dat_gene %>% 
    dplyr::bind_rows(data.frame(SCFA          = dat_gene$SCFA[1],
                                gene          = gen,
                                intervention  = "Baseline median TPM abd.",
                                pvalue_signif = "",
                                effect_label  = case_when(dat_gene$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                          dat_gene$baseline_medianTPMAbd < .001  ~ paste0(round(dat_gene$baseline_medianTPMAbd, 4), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .01   ~ paste0(round(dat_gene$baseline_medianTPMAbd, 3), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .1    ~ paste0(round(dat_gene$baseline_medianTPMAbd, 2), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < 1     ~ paste0(round(dat_gene$baseline_medianTPMAbd, 1), " TPM"),
                                                          TRUE ~ paste0(round(dat_gene$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(gene          = factor(gene, levels = rev(unique(gene))),
         # intervention  = gsub(intervention, pattern = "low",  replacement = "[18, 25)"),
         # intervention  = gsub(intervention, pattern = "high", replacement = "[25, 31]"),
         intervention  = factor(intervention, levels = c("Baseline median TPM abd.",
                                                         "Fresh intervention\n(fiber low)",
                                                         "Fresh intervention\n(fiber high)",
                                                         "Pasteurized intervention\n(fiber low)",
                                                         "Pasteurized intervention\n(fiber high)")),
         pvalue_signif = factor(pvalue_signif, levels = c("not significant",
                                                          "significant pos. effect (fresh intervention)",
                                                          "significant neg. effect (fresh intervention)",
                                                          "significant pos. effect (pasteurized intervention)",
                                                          "significant neg. effect (pasteurized intervention)",
                                                          "")),)

# plot
dat_plot %>% 
  ggplot(aes(x = intervention, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("uncorrected p-value", values = c("not significant" = "gray95",
                                                      "significant pos. effect (fresh intervention)" = "#7cbde9",
                                                      "significant neg. effect (fresh intervention)" = "#1F78B4",
                                                      "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                                      "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(SCFA), scales = "free", space = "free") +
  ggtitle("Intervention effects on SCFA genes") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"))

Joint heatmap of all interaction effects

… of uncorrected p-values

Create the diversity interaction data

… same as for the above plot, but also including species that showed significances in any other interaction

Code

# prepare the base dataset for the plot
dat_signif <- dat_SCFAresults %>% 
  filter((freshInt_divLow_pvalue < 0.05) | (freshInt_divHigh_pvalue < 0.05) |
           (pastInt_divLow_pvalue < 0.05) | (pastInt_divHigh_pvalue < 0.05) |
           (freshInt_ageLow_pvalue < 0.05) | (freshInt_ageHigh_pvalue < 0.05) |
           (pastInt_ageLow_pvalue < 0.05) | (pastInt_ageHigh_pvalue < 0.05) |
           (freshInt_sexM_pvalue < 0.05) | (freshInt_sexW_pvalue < 0.05) |
           (pastInt_sexM_pvalue < 0.05) | (pastInt_sexW_pvalue < 0.05) |
           (freshInt_bmiLow_pvalue < 0.05) | (freshInt_bmiHigh_pvalue < 0.05) |
           (pastInt_bmiLow_pvalue < 0.05) | (pastInt_bmiHigh_pvalue < 0.05) |
           (freshInt_fibLow_pvalue < 0.05) | (freshInt_fibHigh_pvalue < 0.05) |
           (pastInt_fibLow_pvalue < 0.05) | (pastInt_fibHigh_pvalue < 0.05))
dat_plot <- data.frame(gene          = rep(dat_signif$gene,  times = 4),
                       SCFA          = rep(dat_signif$SCFA,  times = 4),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(div. low)", "Fresh intervention\n(div. high)",
                                             "Pasteurized intervention\n(div. low)", "Pasteurized intervention\n(div. high)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_divLow_effect, dat_signif$freshInt_divHigh_effect,
                                         dat_signif$pastInt_divLow_effect,  dat_signif$pastInt_divHigh_effect),
                       pvalue        = c(dat_signif$freshInt_divLow_pvalue, dat_signif$freshInt_divHigh_pvalue,
                                         dat_signif$pastInt_divLow_pvalue,  dat_signif$pastInt_divHigh_pvalue),
                       pastEffect_forSorting = rep(dat_signif$pastInt_divLow_effect, times = 4)) %>% 
  arrange(desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((pvalue < 0.05) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (pvalue < 0.05) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (pvalue < 0.05) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (pvalue < 0.05) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 PP",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 PP",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$gene)), function(gen) {
  dat_gene <- dat_plot %>% filter(gene == gen)
  dat_gene %>% 
    dplyr::bind_rows(data.frame(gene          = gen,
                                intervention  = "Baseline median TPM abd.",
                                SCFA          = dat_gene$SCFA[1],
                                pvalue_signif = "",
                                effect_label  = case_when(dat_gene$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                          dat_gene$baseline_medianTPMAbd < .001  ~ paste0(round(dat_gene$baseline_medianTPMAbd, 4), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .01   ~ paste0(round(dat_gene$baseline_medianTPMAbd, 3), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .1    ~ paste0(round(dat_gene$baseline_medianTPMAbd, 2), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < 1     ~ paste0(round(dat_gene$baseline_medianTPMAbd, 1), " TPM"),
                                                          TRUE ~ paste0(round(dat_gene$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot_divIntp <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(gene         = factor(gene, levels = rev(unique(gene))),
         intervention = factor(intervention, levels = c("Baseline median TPM abd.", "Fresh intervention\n(div. low)", "Fresh intervention\n(div. high)",
                                                        "Pasteurized intervention\n(div. low)", "Pasteurized intervention\n(div. high)")),
         text_highlight = case_when(grepl("Baseline", intervention) & (effect_label == "<0.0001%") ~ "no",
                                    pvalue_signif == "not significant" ~ "no",
                                    TRUE ~ "yes"))

Create the age interaction data

… same as for the above plot, but also including species that showed significances in any other interaction

Code

# prepare the base dataset for the plot
dat_signif <- dat_SCFAresults %>% 
  filter((freshInt_divLow_pvalue < 0.05) | (freshInt_divHigh_pvalue < 0.05) |
           (pastInt_divLow_pvalue < 0.05) | (pastInt_divHigh_pvalue < 0.05) |
           (freshInt_ageLow_pvalue < 0.05) | (freshInt_ageHigh_pvalue < 0.05) |
           (pastInt_ageLow_pvalue < 0.05) | (pastInt_ageHigh_pvalue < 0.05) |
           (freshInt_sexM_pvalue < 0.05) | (freshInt_sexW_pvalue < 0.05) |
           (pastInt_sexM_pvalue < 0.05) | (pastInt_sexW_pvalue < 0.05) |
           (freshInt_bmiLow_pvalue < 0.05) | (freshInt_bmiHigh_pvalue < 0.05) |
           (pastInt_bmiLow_pvalue < 0.05) | (pastInt_bmiHigh_pvalue < 0.05) |
           (freshInt_fibLow_pvalue < 0.05) | (freshInt_fibHigh_pvalue < 0.05) |
           (pastInt_fibLow_pvalue < 0.05) | (pastInt_fibHigh_pvalue < 0.05))
dat_plot <- data.frame(gene          = rep(dat_signif$gene,  times = 4),
                       SCFA          = rep(dat_signif$SCFA,  times = 4),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(age low)", "Fresh intervention\n(age high)",
                                             "Pasteurized intervention\n(age low)", "Pasteurized intervention\n(age high)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_ageLow_effect, dat_signif$freshInt_ageHigh_effect,
                                         dat_signif$pastInt_ageLow_effect,  dat_signif$pastInt_ageHigh_effect),
                       pvalue        = c(dat_signif$freshInt_ageLow_pvalue, dat_signif$freshInt_ageHigh_pvalue,
                                         dat_signif$pastInt_ageLow_pvalue,  dat_signif$pastInt_ageHigh_pvalue),
                       pastEffect_forSorting = rep(dat_signif$pastInt_ageLow_effect, times = 4)) %>% 
  arrange(desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((pvalue < 0.05) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (pvalue < 0.05) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (pvalue < 0.05) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (pvalue < 0.05) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 PP",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 PP",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$gene)), function(gen) {
  dat_gene <- dat_plot %>% filter(gene == gen)
  dat_gene %>% 
    dplyr::bind_rows(data.frame(gene          = gen,
                                intervention  = "Baseline median TPM abd.",
                                SCFA          = dat_gene$SCFA[1],
                                pvalue_signif = "",
                                effect_label  = case_when(dat_gene$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                          dat_gene$baseline_medianTPMAbd < .001  ~ paste0(round(dat_gene$baseline_medianTPMAbd, 4), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .01   ~ paste0(round(dat_gene$baseline_medianTPMAbd, 3), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .1    ~ paste0(round(dat_gene$baseline_medianTPMAbd, 2), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < 1     ~ paste0(round(dat_gene$baseline_medianTPMAbd, 1), " TPM"),
                                                          TRUE ~ paste0(round(dat_gene$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot_ageIntp <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(gene         = factor(gene, levels = rev(unique(gene))),
         intervention = factor(intervention, levels = c("Baseline median TPM abd.", "Fresh intervention\n(age low)", "Fresh intervention\n(age high)",
                                                        "Pasteurized intervention\n(age low)", "Pasteurized intervention\n(age high)")),
         text_highlight = case_when(grepl("Baseline", intervention) & (effect_label == "<0.0001%") ~ "no",
                                    pvalue_signif == "not significant" ~ "no",
                                    TRUE ~ "yes"))

create the sex interaction data

… same as for the above plot, but also including species that showed significances in any other interaction

Code

# prepare the base dataset for the plot
dat_signif <- dat_SCFAresults %>% 
  filter((freshInt_ageLow_pvalue < 0.05) | (freshInt_ageHigh_pvalue < 0.05) |
           (pastInt_ageLow_pvalue < 0.05) | (pastInt_ageHigh_pvalue < 0.05) |
           (freshInt_sexM_pvalue < 0.05) | (freshInt_sexW_pvalue < 0.05) |
           (pastInt_sexM_pvalue < 0.05) | (pastInt_sexW_pvalue < 0.05) |
           (freshInt_bmiLow_pvalue < 0.05) | (freshInt_bmiHigh_pvalue < 0.05) |
           (pastInt_bmiLow_pvalue < 0.05) | (pastInt_bmiHigh_pvalue < 0.05))
dat_plot <- data.frame(gene          = rep(dat_signif$gene,  times = 4),
                       SCFA          = rep(dat_signif$SCFA,  times = 4),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(gender male)", "Fresh intervention\n(gender female)",
                                             "Pasteurized intervention\n(gender male)", "Pasteurized intervention\n(gender female)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_sexM_effect, dat_signif$freshInt_sexW_effect,
                                         dat_signif$pastInt_sexM_effect,  dat_signif$pastInt_sexW_effect),
                       pvalue        = c(dat_signif$freshInt_sexM_pvalue, dat_signif$freshInt_sexW_pvalue,
                                         dat_signif$pastInt_sexM_pvalue,  dat_signif$pastInt_sexW_pvalue),
                       pastEffect_forSorting = rep(dat_signif$pastInt_sexM_effect, times = 4)) %>% 
  arrange(desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((pvalue < 0.05) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (pvalue < 0.05) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (pvalue < 0.05) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (pvalue < 0.05) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 PP",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 PP",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$gene)), function(gen) {
  dat_gene <- dat_plot %>% filter(gene == gen)
  dat_gene %>% 
    dplyr::bind_rows(data.frame(gene          = gen,
                                intervention  = "Baseline median TPM abd.",
                                SCFA          = dat_gene$SCFA[1],
                                pvalue_signif = "",
                                effect_label  = case_when(dat_gene$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                          dat_gene$baseline_medianTPMAbd < .001  ~ paste0(round(dat_gene$baseline_medianTPMAbd, 4), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .01   ~ paste0(round(dat_gene$baseline_medianTPMAbd, 3), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .1    ~ paste0(round(dat_gene$baseline_medianTPMAbd, 2), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < 1     ~ paste0(round(dat_gene$baseline_medianTPMAbd, 1), " TPM"),
                                                          TRUE ~ paste0(round(dat_gene$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot_sexIntp <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(gene         = factor(gene, levels = rev(unique(gene))),
         intervention = factor(intervention, levels = c("Baseline median TPM abd.", "Fresh intervention\n(gender male)", "Fresh intervention\n(gender female)",
                                                        "Pasteurized intervention\n(gender male)", "Pasteurized intervention\n(gender female)")),
         text_highlight = case_when(grepl("Baseline", intervention) & (effect_label == "<0.0001%") ~ "no",
                                    pvalue_signif == "not significant" ~ "no",
                                    TRUE ~ "yes"))

create the BMI interaction data

… same as for the above plot, but also including species that showed significances in any other interaction

Code

# prepare the base dataset for the plot
dat_signif <- dat_SCFAresults %>% 
  filter((freshInt_ageLow_pvalue < 0.05) | (freshInt_ageHigh_pvalue < 0.05) |
           (pastInt_ageLow_pvalue < 0.05) | (pastInt_ageHigh_pvalue < 0.05) |
           (freshInt_sexM_pvalue < 0.05) | (freshInt_sexW_pvalue < 0.05) |
           (pastInt_sexM_pvalue < 0.05) | (pastInt_sexW_pvalue < 0.05) |
           (freshInt_bmiLow_pvalue < 0.05) | (freshInt_bmiHigh_pvalue < 0.05) |
           (pastInt_bmiLow_pvalue < 0.05) | (pastInt_bmiHigh_pvalue < 0.05))
dat_plot <- data.frame(gene          = rep(dat_signif$gene,  times = 4),
                       SCFA          = rep(dat_signif$SCFA,  times = 4),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(BMI low)", "Fresh intervention\n(BMI high)",
                                             "Pasteurized intervention\n(BMI low)", "Pasteurized intervention\n(BMI high)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_bmiLow_effect, dat_signif$freshInt_bmiHigh_effect,
                                         dat_signif$pastInt_bmiLow_effect,  dat_signif$pastInt_bmiHigh_effect),
                       pvalue        = c(dat_signif$freshInt_bmiLow_pvalue, dat_signif$freshInt_bmiHigh_pvalue,
                                         dat_signif$pastInt_bmiLow_pvalue,  dat_signif$pastInt_bmiHigh_pvalue),
                       pastEffect_forSorting = rep(dat_signif$pastInt_bmiLow_effect, times = 4)) %>% 
  arrange(desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((pvalue < 0.05) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (pvalue < 0.05) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (pvalue < 0.05) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (pvalue < 0.05) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 PP",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 PP",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$gene)), function(gen) {
  dat_gene <- dat_plot %>% filter(gene == gen)
  dat_gene %>% 
    dplyr::bind_rows(data.frame(gene          = gen,
                                intervention  = "Baseline median TPM abd.",
                                SCFA          = dat_gene$SCFA[1],
                                pvalue_signif = "",
                                effect_label  = case_when(dat_gene$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                          dat_gene$baseline_medianTPMAbd < .001  ~ paste0(round(dat_gene$baseline_medianTPMAbd, 4), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .01   ~ paste0(round(dat_gene$baseline_medianTPMAbd, 3), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .1    ~ paste0(round(dat_gene$baseline_medianTPMAbd, 2), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < 1     ~ paste0(round(dat_gene$baseline_medianTPMAbd, 1), " TPM"),
                                                          TRUE ~ paste0(round(dat_gene$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot_bmiIntp <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(gene         = factor(gene, levels = rev(unique(gene))),
         intervention = factor(intervention, levels = c("Baseline median TPM abd.", "Fresh intervention\n(BMI low)", "Fresh intervention\n(BMI high)",
                                                        "Pasteurized intervention\n(BMI low)", "Pasteurized intervention\n(BMI high)")),
         text_highlight = case_when(grepl("Baseline", intervention) & (effect_label == "<0.0001%") ~ "no",
                                    pvalue_signif == "not significant" ~ "no",
                                    TRUE ~ "yes"))

Create the fiber interaction data

… same as for the above plot, but also including species that showed significances in any other interaction

Code

# prepare the base dataset for the plot
dat_signif <- dat_SCFAresults %>% 
  filter((freshInt_divLow_pvalue < 0.05) | (freshInt_divHigh_pvalue < 0.05) |
           (pastInt_divLow_pvalue < 0.05) | (pastInt_divHigh_pvalue < 0.05) |
           (freshInt_ageLow_pvalue < 0.05) | (freshInt_ageHigh_pvalue < 0.05) |
           (pastInt_ageLow_pvalue < 0.05) | (pastInt_ageHigh_pvalue < 0.05) |
           (freshInt_sexM_pvalue < 0.05) | (freshInt_sexW_pvalue < 0.05) |
           (pastInt_sexM_pvalue < 0.05) | (pastInt_sexW_pvalue < 0.05) |
           (freshInt_bmiLow_pvalue < 0.05) | (freshInt_bmiHigh_pvalue < 0.05) |
           (pastInt_bmiLow_pvalue < 0.05) | (pastInt_bmiHigh_pvalue < 0.05) |
           (freshInt_fibLow_pvalue < 0.05) | (freshInt_fibHigh_pvalue < 0.05) |
           (pastInt_fibLow_pvalue < 0.05) | (pastInt_fibHigh_pvalue < 0.05))
dat_plot <- data.frame(gene          = rep(dat_signif$gene,  times = 4),
                       SCFA          = rep(dat_signif$SCFA,  times = 4),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(fiber low)", "Fresh intervention\n(fiber high)",
                                             "Pasteurized intervention\n(fiber low)", "Pasteurized intervention\n(fiber high)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_fibLow_effect, dat_signif$freshInt_fibHigh_effect,
                                         dat_signif$pastInt_fibLow_effect,  dat_signif$pastInt_fibHigh_effect),
                       pvalue        = c(dat_signif$freshInt_fibLow_pvalue, dat_signif$freshInt_fibHigh_pvalue,
                                         dat_signif$pastInt_fibLow_pvalue,  dat_signif$pastInt_fibHigh_pvalue),
                       pastEffect_forSorting = rep(dat_signif$pastInt_fibLow_effect, times = 4)) %>% 
  arrange(desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((pvalue < 0.05) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (pvalue < 0.05) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (pvalue < 0.05) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (pvalue < 0.05) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 PP",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " PP"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 PP",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$gene)), function(gen) {
  dat_gene <- dat_plot %>% filter(gene == gen)
  dat_gene %>% 
    dplyr::bind_rows(data.frame(gene          = gen,
                                intervention  = "Baseline median TPM abd.",
                                SCFA          = dat_gene$SCFA[1],
                                pvalue_signif = "",
                                effect_label  = case_when(dat_gene$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                          dat_gene$baseline_medianTPMAbd < .001  ~ paste0(round(dat_gene$baseline_medianTPMAbd, 4), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .01   ~ paste0(round(dat_gene$baseline_medianTPMAbd, 3), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < .1    ~ paste0(round(dat_gene$baseline_medianTPMAbd, 2), " TPM"),
                                                          dat_gene$baseline_medianTPMAbd < 1     ~ paste0(round(dat_gene$baseline_medianTPMAbd, 1), " TPM"),
                                                          TRUE ~ paste0(round(dat_gene$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot_fibIntp <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(gene         = factor(gene, levels = rev(unique(gene))),
         intervention = factor(intervention, levels = c("Baseline median TPM abd.", "Fresh intervention\n(fiber low)", "Fresh intervention\n(fiber high)",
                                                        "Pasteurized intervention\n(fiber low)", "Pasteurized intervention\n(fiber high)")),
         text_highlight = case_when(grepl("Baseline", intervention) & (effect_label == "<0.0001%") ~ "no",
                                    pvalue_signif == "not significant" ~ "no",
                                    TRUE ~ "yes"))

create the joint plot

Code

# plots
gg_heatmap_divInt <- dat_plot_divIntp %>% 
  ggplot(aes(x = intervention, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("uncorrected p-value", values = c("not significant" = "gray95",
                                                      "significant pos. effect (fresh intervention)" = "#7cbde9",
                                                      "significant neg. effect (fresh intervention)" = "#1F78B4",
                                                      "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                                      "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(SCFA), scales = "free", space = "free") +
  ggtitle("Intervention effects on species") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.text = element_blank())
gg_heatmap_ageInt <- dat_plot_ageIntp %>% 
  filter(!grepl("Baseline", intervention)) %>% 
  ggplot(aes(x = intervention, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("uncorrected p-value", values = c("not significant" = "gray95",
                                                      "significant pos. effect (fresh intervention)" = "#7cbde9",
                                                      "significant neg. effect (fresh intervention)" = "#1F78B4",
                                                      "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                                      "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(SCFA), scales = "free", space = "free") +
  ggtitle("Intervention effects on species") +
  theme(axis.title  = element_blank(),
        axis.text.y = element_blank(),
        panel.grid  = element_blank(),
        strip.text  = element_blank())
gg_heatmap_sexInt <- dat_plot_sexIntp %>% 
  filter(!grepl("Baseline", intervention)) %>% 
  ggplot(aes(x = intervention, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("uncorrected p-value", values = c("not significant" = "gray95",
                                                      "significant pos. effect (fresh intervention)" = "#7cbde9",
                                                      "significant neg. effect (fresh intervention)" = "#1F78B4",
                                                      "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                                      "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(SCFA), scales = "free", space = "free") +
  ggtitle("Intervention effects on species") +
  theme(axis.title  = element_blank(),
        axis.text.y = element_blank(),
        panel.grid  = element_blank(),
        strip.text  = element_blank())
gg_heatmap_bmiInt <- dat_plot_bmiIntp %>% 
  filter(!grepl("Baseline", intervention)) %>% 
  ggplot(aes(x = intervention, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("uncorrected p-value", values = c("not significant" = "gray95",
                                                      "significant pos. effect (fresh intervention)" = "#7cbde9",
                                                      "significant neg. effect (fresh intervention)" = "#1F78B4",
                                                      "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                                      "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  ggtitle("Intervention effects on species") +
  facet_grid(rows = vars(SCFA), scales = "free", space = "free") +
  theme(axis.title  = element_blank(),
        axis.text.y = element_blank(),
        panel.grid  = element_blank())
gg_heatmap_fibInt <- dat_plot_fibIntp %>% 
  filter(!grepl("Baseline", intervention)) %>% 
  ggplot(aes(x = intervention, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("uncorrected p-value", values = c("not significant" = "gray95",
                                                      "significant pos. effect (fresh intervention)" = "#7cbde9",
                                                      "significant neg. effect (fresh intervention)" = "#1F78B4",
                                                      "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                                      "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  ggtitle("Intervention effects on species") +
  facet_grid(rows = vars(SCFA), scales = "free", space = "free") +
  theme(axis.title  = element_blank(),
        axis.text.y = element_blank(),
        panel.grid  = element_blank())

# align the plots
(gg_heatmap_divInt + gg_heatmap_ageInt + gg_heatmap_sexInt + gg_heatmap_bmiInt + gg_heatmap_fibInt) +
  patchwork::plot_layout(widths = c(.36, .32, .32, .32, .32), nrow = 1, guides = "collect")

Code

# ggsave("functional_geneScreening_allInteractions_pvalues.png", width = 30, height = 30)

Joint plot of condensed information

Focus on the four biggest phylum groups: Firmicutes, Bacteroidetes, Actinobacteria and Proteobacteria

Code

# data preparation
plot_dat_wide <- dat_SCFAresults %>% 
  group_by(SCFA) %>% 
  dplyr::summarize(n_genes                      = n(),
                   n_freshSignifEffects_overall = sum(freshInt_pvalue         < 0.05),
                   n_freshSignifEffects_divLow  = sum(freshInt_divLow_pvalue  < 0.05),
                   n_freshSignifEffects_divHigh = sum(freshInt_divHigh_pvalue < 0.05),
                   n_freshSignifEffects_ageLow  = sum(freshInt_ageLow_pvalue  < 0.05),
                   n_freshSignifEffects_ageHigh = sum(freshInt_ageHigh_pvalue < 0.05),
                   n_freshSignifEffects_sexM    = sum(freshInt_sexM_pvalue    < 0.05),
                   n_freshSignifEffects_sexW    = sum(freshInt_sexW_pvalue    < 0.05),
                   n_freshSignifEffects_bmiLow  = sum(freshInt_bmiLow_pvalue  < 0.05),
                   n_freshSignifEffects_bmiHigh = sum(freshInt_bmiHigh_pvalue < 0.05),
                   n_freshSignifEffects_fibLow  = sum(freshInt_fibLow_pvalue  < 0.05),
                   n_freshSignifEffects_fibHigh = sum(freshInt_fibHigh_pvalue < 0.05),
                   n_pastSignifEffects_overall  = sum(pastInt_pvalue          < 0.05),
                   n_pastSignifEffects_divLow   = sum(pastInt_divLow_pvalue   < 0.05),
                   n_pastSignifEffects_divHigh  = sum(pastInt_divHigh_pvalue  < 0.05),
                   n_pastSignifEffects_ageLow   = sum(pastInt_ageLow_pvalue   < 0.05),
                   n_pastSignifEffects_ageHigh  = sum(pastInt_ageHigh_pvalue  < 0.05),
                   n_pastSignifEffects_sexM     = sum(pastInt_sexM_pvalue     < 0.05),
                   n_pastSignifEffects_sexW     = sum(pastInt_sexW_pvalue     < 0.05),
                   n_pastSignifEffects_bmiLow   = sum(pastInt_bmiLow_pvalue   < 0.05),
                   n_pastSignifEffects_bmiHigh  = sum(pastInt_bmiHigh_pvalue  < 0.05),
                   n_pastSignifEffects_fibLow   = sum(pastInt_fibLow_pvalue   < 0.05),
                   n_pastSignifEffects_fibHigh  = sum(pastInt_fibHigh_pvalue  < 0.05))


# reshape dataset to long format
plot_dat_long <- data.frame(SCFA                 = rep(plot_dat_wide$SCFA, times = sum(grepl("freshSignifEffects", colnames(plot_dat_wide)))),
                            n_genes              = rep(plot_dat_wide$n_genes, times = sum(grepl("freshSignifEffects", colnames(plot_dat_wide)))),
                            effect_group         = rep(c("overall", "divInt", "divInt", "ageInt", "ageInt", "sexInt", "sexInt", "bmiInt", "bmiInt", "fibInt", "fibInt"),
                                                       each = nrow(plot_dat_wide)),
                            effect_type          = rep(c("overall", "divLow", "divHigh", "ageLow", "ageHigh", "sexM", "sexW", "bmiLow", "bmiHigh", "fibLow", "fibHigh"),
                                                       each = nrow(plot_dat_wide)),
                            n_freshSignifEffects = c(plot_dat_wide$n_freshSignifEffects_overall,
                                                     plot_dat_wide$n_freshSignifEffects_divLow,
                                                     plot_dat_wide$n_freshSignifEffects_divHigh,
                                                     plot_dat_wide$n_freshSignifEffects_ageLow,
                                                     plot_dat_wide$n_freshSignifEffects_ageHigh,
                                                     plot_dat_wide$n_freshSignifEffects_sexM,
                                                     plot_dat_wide$n_freshSignifEffects_sexW,
                                                     plot_dat_wide$n_freshSignifEffects_bmiLow,
                                                     plot_dat_wide$n_freshSignifEffects_bmiHigh,
                                                     plot_dat_wide$n_freshSignifEffects_fibLow,
                                                     plot_dat_wide$n_freshSignifEffects_fibHigh),
                            n_pastSignifEffects  = c(plot_dat_wide$n_pastSignifEffects_overall,
                                                     plot_dat_wide$n_pastSignifEffects_divLow,
                                                     plot_dat_wide$n_pastSignifEffects_divHigh,
                                                     plot_dat_wide$n_pastSignifEffects_ageLow,
                                                     plot_dat_wide$n_pastSignifEffects_ageHigh,
                                                     plot_dat_wide$n_pastSignifEffects_sexM,
                                                     plot_dat_wide$n_pastSignifEffects_sexW,
                                                     plot_dat_wide$n_pastSignifEffects_bmiLow,
                                                     plot_dat_wide$n_pastSignifEffects_bmiHigh,
                                                     plot_dat_wide$n_pastSignifEffects_fibLow,
                                                     plot_dat_wide$n_pastSignifEffects_fibHigh)) %>% 
  mutate(SCFA         = gsub(SCFA, pattern = "succinate",  replacement = "Succinate"),
         SCFA         = gsub(SCFA, pattern = "propionate", replacement = "Propionate"),
         SCFA_label   = paste0(SCFA, "\n", n_genes, " genes"),
         effect_group = factor(effect_group, levels = c("overall", "divInt", "ageInt", "sexInt", "bmiInt", "fibInt")),
         effect_type  = factor(effect_type, levels = rev(c("overall", "divLow", "divHigh", "ageLow", "ageHigh", "sexM", "sexW", "bmiLow", "bmiHigh", "fibLow", "fibHigh"))))

# make the dataset even longer
plot_dat_longer <- data.frame(SCFA         = rep(plot_dat_long$SCFA,         times = 2),
                              n_genes      = rep(plot_dat_long$n_genes,      times = 2),
                              SCFA_label   = rep(plot_dat_long$SCFA_label,   times = 2),
                              effect_group = rep(plot_dat_long$effect_group, times = 2),
                              effect_type  = rep(plot_dat_long$effect_type,  times = 2),
                              intervention = rep(c("Fresh", "Past."), each = nrow(plot_dat_long)),
                              parameter    = "n_signifEffects",
                              value        = c(plot_dat_long$n_freshSignifEffects,
                                               plot_dat_long$n_pastSignifEffects)) %>% 
  group_by(SCFA, parameter, effect_group, intervention) %>% 
  mutate(alpha_value = case_when(effect_group == "overall"  ~ 0.8,
                                 value == min(value)        ~ 0,
                                 TRUE                       ~ 1)) %>% 
  ungroup() %>% 
  mutate(fill_tiles = case_when(effect_type == "overall" ~ "no",
                                TRUE ~ SCFA),
         alpha_tiles = case_when(effect_type %in% c("divLow", "ageLow", "sexM", "bmiLow", "fibInt") ~ 0.05,
                                 TRUE ~ 0.15))
plot_dat_longer <- plot_dat_longer %>% mutate(SCFA_label = factor(SCFA_label, levels = plot_dat_longer %>% arrange(desc(n_genes)) %>% 
                                                                    pull(SCFA_label) %>% unique()))


# create a color vector for color coding the interaction terms
col_vector <- RColorBrewer::brewer.pal(6, name = "Dark2")

# plot
ggplot(plot_dat_longer, aes(x = intervention, y = effect_type)) +
  geom_tile(aes(fill = fill_tiles, alpha = alpha_tiles)) +
  geom_text(aes(label = value, alpha = alpha_value)) +
  facet_grid(effect_group ~ SCFA_label, scales = "free") +
  scale_fill_manual(values = c("no"             = "white",
                               "Pyruvate"       = col_vector[1],
                               "Acetyl-CoA"     = col_vector[2],
                               "Acetate"        = col_vector[3],
                               "succinate"      = col_vector[4],
                               "Butyryl-CoA"    = col_vector[5],
                               "propionate"     = col_vector[6])) +
  theme(axis.title      = element_blank(),
        panel.grid      = element_blank(),
        legend.position = "none",
        strip.background.x = element_rect(fill = "gray97", color = "gray97"),
        strip.text.y    = element_blank())

Code

# ggsave("functional_geneScreening_allInteractions_table.png", width = 7, height = 3.5)

Main joint SCFA gene figure for the publication

Code

# 1) plot of overall effects
gg_overall <- dat_plot_overall %>% 
  mutate(pvalue_signif = gsub(pvalue_signif, pattern = "significant ",  replacement = ""),
         pvalue_signif = gsub(pvalue_signif, pattern = " intervention", replacement = ""),
         pvalue_signif = gsub(pvalue_signif, pattern = "pasteurized",   replacement = "past."),
         pvalue_signif = factor(pvalue_signif, levels = c("not significant", "pos. effect (fresh)",
                                                          "neg. effect (fresh)", "pos. effect (past.)",
                                                          "neg. effect (past.)", "")),
         effect_label  = gsub(effect_label, pattern = "\n", replacement = " ")) %>% 
  ggplot(aes(x = intervention, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label, size = text_size)) +
  scale_fill_manual("q-value", values = c("not significant" = "gray95",
                                          "pos. effect (fresh)" = "#7cbde9",
                                          "neg. effect (fresh)" = "#1F78B4",
                                          "pos. effect (past.)" = "#ffb366",
                                          "neg. effect (past.)" = "#FF7F00"),
                    na.value = "white") +
  scale_size(range = c(3.5, 4), guide = "none") +
  theme_minimal(base_size = 15) +
  theme(plot.title = element_text(hjust = 0.5),
        axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"),
        legend.position = "none")

# 1.5) create the legend separately, also to ensure that all color categories are plotted
category_labels <- c("not significant","pos. effect (fresh)","neg. effect (fresh)",
                     "pos. effect (past.)","neg. effect (past.)")
gg_base    <- data.frame(x       = rnorm(n = 5),
                         y       = rnorm(n = 5),
                         col_var = factor(category_labels, levels = category_labels)) %>% 
  ggplot(aes(x = x, y = y, fill = col_var)) +
  geom_tile() +
  scale_fill_manual("q-value", values = c("not significant"     = "gray95",
                                          "pos. effect (fresh)" = "#7cbde9",
                                          "neg. effect (fresh)" = "#1F78B4",
                                          "pos. effect (past.)" = "#ffb366",
                                          "neg. effect (past.)" = "#FF7F00"),
                    na.value = "white") +
  theme_minimal(base_size = 15) +
  theme(legend.position = "bottom")
gg_legend  <- ggpubr::get_legend(gg_base) %>% ggpubr::as_ggplot()


# 2) plot of diversity-specific effects
# NOT necessary since no effect is significant
# gg_div <- dat_plot_divInt %>% 
#   filter(intervention != "Baseline median TPM abd.") %>% 
#   mutate(pvalue_signif = gsub(pvalue_signif, pattern = "significant ",  replacement = ""),
#          pvalue_signif = gsub(pvalue_signif, pattern = " intervention", replacement = ""),
#          pvalue_signif = gsub(pvalue_signif, pattern = "pasteurized",   replacement = "past."),
#          pvalue_signif = factor(pvalue_signif, levels = c("not significant", "pos. effect (fresh)",
#                                                           "neg. effect (fresh)", "pos. effect (past.)",
#                                                           "neg. effect (past.)", ""))) %>% 
#   mutate(intervention2 = gsub(intervention,  pattern = "Fresh intervention\n\\(",       replacement = ""),
#          intervention2 = gsub(intervention2, pattern = "Pasteurized intervention\n\\(", replacement = ""),
#          intervention2 = gsub(intervention2, pattern = "\\)", replacement = ""),
#          intervention2 = gsub(intervention2, pattern = "div. low",  replacement = "div. [2.8, 3.85)"),
#          intervention2 = gsub(intervention2, pattern = "div. high", replacement = "div. [3.85, 4.4)"),
#          intervention3 = gsub(intervention,  pattern = "\n\\(div. low\\)",  replacement = ""),
#          intervention3 = gsub(intervention3, pattern = "\n\\(div. high\\)", replacement = "")) %>%
#   ggplot(aes(x = intervention2, y = gene)) +
#   geom_tile(aes(fill = pvalue_signif)) +
#   geom_text(aes(label = effect_label)) +
#   scale_fill_manual("q-value", values = c("not significant" = "gray95",
#                                           "pos. effect (fresh)" = "#7cbde9",
#                                           "neg. effect (fresh)" = "#1F78B4",
#                                           "pos. effect (past.)" = "#ffb366",
#                                           "neg. effect (past.)" = "#FF7F00"),
#                     na.value = "white") +
#   guides(fill = guide_legend(nrow = 1)) +
#   facet_grid(cols = vars(intervention3), scales = "free", space = "free") +
#   theme_minimal(base_size = 15) +
#   theme(plot.title = element_text(hjust = 0.5),
#         axis.title = element_blank(),
#         panel.grid = element_blank(),
#         strip.text.x     = element_blank(),
#         strip.background = element_rect(fill = "gray97", color = "gray97"),
#         legend.position  = "none")


# 3) plot of age-specific effects
gg_age <- dat_plot_ageInt %>% 
  filter(intervention != "Baseline median TPM abd.") %>% 
  mutate(pvalue_signif = gsub(pvalue_signif, pattern = "significant ",  replacement = ""),
         pvalue_signif = gsub(pvalue_signif, pattern = " intervention", replacement = ""),
         pvalue_signif = gsub(pvalue_signif, pattern = "pasteurized",   replacement = "past."),
         pvalue_signif = factor(pvalue_signif, levels = c("not significant", "pos. effect (fresh)",
                                                          "neg. effect (fresh)", "pos. effect (past.)",
                                                          "neg. effect (past.)", ""))) %>% 
  mutate(intervention2 = gsub(intervention,  pattern = "Fresh intervention\n\\(",       replacement = ""),
         intervention2 = gsub(intervention2, pattern = "Pasteurized intervention\n\\(", replacement = ""),
         intervention2 = gsub(intervention2, pattern = "\\)", replacement = ""),
         intervention2 = gsub(intervention2, pattern = "age low",  replacement = "age 21-49"),
         intervention2 = gsub(intervention2, pattern = "age high", replacement = "age 50-69"),
         intervention3 = gsub(intervention,  pattern = "\n\\(age low\\)",  replacement = ""),
         intervention3 = gsub(intervention3, pattern = "\n\\(age high\\)", replacement = "")) %>%
  ggplot(aes(x = intervention2, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("q-value", values = c("not significant" = "gray95",
                                          "pos. effect (fresh)" = "#7cbde9",
                                          "neg. effect (fresh)" = "#1F78B4",
                                          "pos. effect (past.)" = "#ffb366",
                                          "neg. effect (past.)" = "#FF7F00"),
                    na.value = "white") +
  guides(fill = guide_legend(nrow = 1)) +
  facet_grid(cols = vars(intervention3), scales = "free", space = "free") +
  theme_minimal(base_size = 15) +
  theme(plot.title = element_text(hjust = 0.5),
        axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.text.x     = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"),
        legend.position  = "none")


# 4) plot of gender-specific effects
gg_sex <- dat_plot_sexInt %>% 
  filter(intervention != "Baseline median TPM abd.") %>% 
  mutate(pvalue_signif = gsub(pvalue_signif, pattern = "significant ",  replacement = ""),
         pvalue_signif = gsub(pvalue_signif, pattern = " intervention", replacement = ""),
         pvalue_signif = gsub(pvalue_signif, pattern = "pasteurized",   replacement = "past."),
         pvalue_signif = factor(pvalue_signif, levels = c("not significant", "pos. effect (fresh)",
                                                          "neg. effect (fresh)", "pos. effect (past.)",
                                                          "neg. effect (past.)", ""))) %>% 
  mutate(intervention2 = gsub(intervention,  pattern = "Fresh intervention\n\\(",       replacement = ""),
         intervention2 = gsub(intervention2, pattern = "Pasteurized intervention\n\\(", replacement = ""),
         intervention2 = gsub(intervention2, pattern = "\\)", replacement = ""),
         intervention2 = gsub(intervention2, pattern = "gender ",  replacement = ""),
         intervention2 = factor(intervention2, levels = c("male", "female")),
         intervention3 = gsub(intervention,  pattern = "\n\\(gender male\\)",  replacement = ""),
         intervention3 = gsub(intervention3, pattern = "\n\\(gender female\\)", replacement = "")) %>%
  ggplot(aes(x = intervention2, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("q-value", values = c("not significant" = "gray95",
                                          "pos. effect (fresh)" = "#7cbde9",
                                          "neg. effect (fresh)" = "#1F78B4",
                                          "pos. effect (past.)" = "#ffb366",
                                          "neg. effect (past.)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(cols = vars(intervention3), scales = "free", space = "free") +
  theme_minimal(base_size = 15) +
  theme(plot.title = element_text(hjust = 0.5),
        axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.text.x     = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"),
        legend.position = "none")

# 5) plot of BMI-specific effects
gg_bmi <- dat_plot_bmiInt %>% 
  filter(intervention != "Baseline median TPM abd.") %>% 
  mutate(pvalue_signif = gsub(pvalue_signif, pattern = "significant ",  replacement = ""),
         pvalue_signif = gsub(pvalue_signif, pattern = " intervention", replacement = ""),
         pvalue_signif = gsub(pvalue_signif, pattern = "pasteurized",   replacement = "past."),
         pvalue_signif = factor(pvalue_signif, levels = c("not significant", "pos. effect (fresh)",
                                                          "neg. effect (fresh)", "pos. effect (past.)",
                                                          "neg. effect (past.)", ""))) %>% 
  mutate(intervention2 = gsub(intervention,  pattern = "Fresh intervention\n\\(",       replacement = ""),
         intervention2 = gsub(intervention2, pattern = "Pasteurized intervention\n\\(", replacement = ""),
         intervention2 = gsub(intervention2, pattern = "\\)", replacement = ""),
         intervention2 = gsub(intervention2, pattern = "BMI low",  replacement = "BMI [18, 25)"),
         intervention2 = gsub(intervention2, pattern = "BMI high", replacement = "BMI [25, 31]"),
         intervention3 = gsub(intervention,  pattern = "\n\\(BMI low\\)",  replacement = ""),
         intervention3 = gsub(intervention3, pattern = "\n\\(BMI high\\)", replacement = "")) %>%
  ggplot(aes(x = intervention2, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("q-value", values = c("not significant" = "gray95",
                                          "pos. effect (fresh)" = "#7cbde9",
                                          "neg. effect (fresh)" = "#1F78B4",
                                          "pos. effect (past.)" = "#ffb366",
                                          "neg. effect (past.)" = "#FF7F00"),
                    na.value = "white") +
  # facet_grid(rows = vars(phylum), cols = vars(intervention3), scales = "free", space = "free") +
  facet_grid(cols = vars(intervention3), scales = "free", space = "free") +
  # ggtitle("BMI-specific effects") +
  theme_minimal(base_size = 15) +
  theme(plot.title = element_text(hjust = 0.5),
        axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.text.x     = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"),
        legend.position = "none")


# 6) plot of fiber-specific effects
gg_fib <- dat_plot_fibInt %>% 
  filter(intervention != "Baseline median TPM abd.") %>% 
  mutate(pvalue_signif = gsub(pvalue_signif, pattern = "significant ",  replacement = ""),
         pvalue_signif = gsub(pvalue_signif, pattern = " intervention", replacement = ""),
         pvalue_signif = gsub(pvalue_signif, pattern = "pasteurized",   replacement = "past."),
         pvalue_signif = factor(pvalue_signif, levels = c("not significant", "pos. effect (fresh)",
                                                          "neg. effect (fresh)", "pos. effect (past.)",
                                                          "neg. effect (past.)", ""))) %>% 
  mutate(intervention2 = gsub(intervention,  pattern = "Fresh intervention\n\\(",       replacement = ""),
         intervention2 = gsub(intervention2, pattern = "Pasteurized intervention\n\\(", replacement = ""),
         intervention2 = gsub(intervention2, pattern = "\\)", replacement = ""),
         intervention2 = gsub(intervention2, pattern = "fiber low",  replacement = "fiber (8, 30)"),
         intervention2 = gsub(intervention2, pattern = "fiber high", replacement = "fiber [30, 58)"),
         intervention3 = gsub(intervention,  pattern = "\n\\(fiber low\\)",  replacement = ""),
         intervention3 = gsub(intervention3, pattern = "\n\\(fiber high\\)", replacement = "")) %>%
  ggplot(aes(x = intervention2, y = gene)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("q-value", values = c("not significant" = "gray95",
                                          "pos. effect (fresh)" = "#7cbde9",
                                          "neg. effect (fresh)" = "#1F78B4",
                                          "pos. effect (past.)" = "#ffb366",
                                          "neg. effect (past.)" = "#FF7F00"),
                    na.value = "white") +
  guides(fill = guide_legend(nrow = 1)) +
  facet_grid(cols = vars(intervention3), scales = "free", space = "free") +
  theme_minimal(base_size = 15) +
  theme(plot.title = element_text(hjust = 0.5),
        axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.text.x     = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"),
        legend.position  = "none")


# 7) plot of the overview table of the uncorrected p-value tendencies
gg_table <- plot_dat_longer %>% 
  mutate(effect_type = case_when(effect_type == "divLow"  ~ "div. [2.8, 3.85)",
                                 effect_type == "divHigh" ~ "div. [3.85, 4.4)",
                                 effect_type == "ageLow"  ~ "age 21-49",
                                 effect_type == "ageHigh" ~ "age 50-69",
                                 effect_type == "sexM"    ~ "male",
                                 effect_type == "sexW"    ~ "female",
                                 effect_type == "bmiLow"  ~ "BMI [18, 25)",
                                 effect_type == "bmiHigh" ~ "BMI [25, 31]",
                                 effect_type == "fibLow"  ~ "fiber (8, 30)",
                                 effect_type == "fibHigh" ~ "fiber [30, 58)",
                                 TRUE                     ~ effect_type),
         effect_type = factor(effect_type, levels = c("overall",
                                                      "div. [2.8, 3.85)", "div. [3.85, 4.4)",
                                                      "age 50-69", "age 21-49",
                                                      "female", "male",
                                                      "BMI [25, 31]", "BMI [18, 25)",
                                                      "fiber (8, 30)", "fiber [30, 58)"))) %>% 
  ggplot(aes(x = intervention, y = effect_type)) +
  geom_tile(aes(fill = fill_tiles, alpha = alpha_tiles)) +
  geom_text(aes(label = value, alpha = alpha_value)) +
  facet_grid(effect_group ~ SCFA_label, scales = "free") +
  scale_fill_manual(values = c("no"             = "white",
                               "Pyruvate"       = col_vector[1],
                               "Acetyl-CoA"     = col_vector[2],
                               "Acetate"        = col_vector[3],
                               "Succinate"      = col_vector[4],
                               "Butyryl-CoA"    = col_vector[5],
                               "Propionate"     = col_vector[6])) +
  theme_minimal(base_size = 14) +
  theme(plot.title      = element_text(hjust = 0.5),
        axis.title      = element_blank(),
        panel.grid      = element_blank(),
        legend.position = "none",
        strip.background.x = element_rect(fill = "gray97", color = "gray97"),
        strip.text.y    = element_blank())


# 8) small plot just for adding some in between titles to the joint plot
text_dat <- data.frame(x     = 0.5,
                       y     = 0.6,
                       label = c("Overall effects on SCFA gene abundance", "Stratified effects"))
gg_header_base <- ggplot() +
  geom_hline(yintercept = 0) +
  xlim(c(0,1)) + ylim(c(0,1)) +
  theme_minimal(base_size = 15) +
  theme(panel.grid        = element_blank(),
        plot.tag.location = "plot",
        plot.tag          = element_text(size = 18, margin = margin(t = -5)),
        axis.text         = element_blank(),
        axis.title        = element_blank(),
        plot.margin       = margin(t = 0, b = 0, l = 0, r = 0),
        legend.position   = "none")
gg_headerA <- gg_header_base +
  geom_text(data = text_dat %>% dplyr::slice(1), aes(x = x, y = y, label = label), size = 5) +
  labs(tag = "(A)")
gg_headerB <- gg_header_base +
  geom_text(data = text_dat %>% dplyr::slice(2), aes(x = x, y = y, label = label), size = 5) +
  labs(tag = "(B)")

# 9) joint plot
(gg_headerA + gg_overall + gg_legend + gg_headerB + gg_age + gg_sex + gg_bmi + gg_fib) +
  patchwork::plot_layout(ncol = 1, heights = c(.2,1.1,.9,.2,.8,.6,.6,1.3))

Code

# ggsave("FigureS15_geneScreening.png", width = 13, height = 10)

additional summary table of trends

Code

gg_table

KO Results (genes)

Available information

Code

dat_KOresults %>% 
  head(5) %>% 
  kable() %>% 
  kable_styling()

SCFA	gene	relFreq_zeroTPM	relFreq_min10TPM	min_tpmAbd	mean_tpmAbd	median_tpmAbd	max_tpmAbd	baseline_medianTPMAbd	baseline_prevalence	postFresh_medianTPMAbd	postFresh_prevalence	postPast_medianTPMAbd	postPast_prevalence	model	model_deviance	model_meanAbsResid	freshInt_effect	freshInt_se	freshInt_pvalue	pastInt_effect	pastInt_se	pastInt_pvalue	phaseEffect	genderEffect	ageEffect	bmiEffect	freshInt_ageLow_effect	freshInt_ageHigh_effect	freshInt_ageLow_se	freshInt_ageHigh_se	freshInt_ageLow_pvalue	freshInt_ageHigh_pvalue	freshInt_sexM_effect	freshInt_sexW_effect	freshInt_sexM_se	freshInt_sexW_se	freshInt_sexM_pvalue	freshInt_sexW_pvalue	freshInt_bmiLow_effect	freshInt_bmiHigh_effect	freshInt_bmiLow_se	freshInt_bmiHigh_se	freshInt_bmiLow_pvalue	freshInt_bmiHigh_pvalue	freshInt_fibLow_effect	freshInt_fibHigh_effect	freshInt_fibLow_se	freshInt_fibHigh_se	freshInt_fibLow_pvalue	freshInt_fibHigh_pvalue	freshInt_divLow_effect	freshInt_divHigh_effect	freshInt_divLow_se	freshInt_divHigh_se	freshInt_divLow_pvalue	freshInt_divHigh_pvalue	pastInt_ageLow_effect	pastInt_ageHigh_effect	pastInt_ageLow_se	pastInt_ageHigh_se	pastInt_ageLow_pvalue	pastInt_ageHigh_pvalue	pastInt_sexM_effect	pastInt_sexW_effect	pastInt_sexM_se	pastInt_sexW_se	pastInt_sexM_pvalue	pastInt_sexW_pvalue	pastInt_bmiLow_effect	pastInt_bmiHigh_effect	pastInt_bmiLow_se	pastInt_bmiHigh_se	pastInt_bmiLow_pvalue	pastInt_bmiHigh_pvalue	pastInt_fibLow_effect	pastInt_fibHigh_effect	pastInt_fibLow_se	pastInt_fibHigh_se	pastInt_fibLow_pvalue	pastInt_fibHigh_pvalue	pastInt_divLow_effect	pastInt_divHigh_effect	pastInt_divLow_se	pastInt_divHigh_se	pastInt_divLow_pvalue	pastInt_divHigh_pvalue	freshInt_pvalue_correct	pastInt_pvalue_correct	freshInt_ageLow_pvalue_correct	pastInt_ageLow_pvalue_correct	freshInt_ageHigh_pvalue_correct	pastInt_ageHigh_pvalue_correct	freshInt_sexM_pvalue_correct	pastInt_sexM_pvalue_correct	freshInt_sexW_pvalue_correct	pastInt_sexW_pvalue_correct	freshInt_bmiLow_pvalue_correct	pastInt_bmiLow_pvalue_correct	freshInt_bmiHigh_pvalue_correct	pastInt_bmiHigh_pvalue_correct	freshInt_fibLow_pvalue_correct	pastInt_fibLow_pvalue_correct	freshInt_fibHigh_pvalue_correct	pastInt_fibHigh_pvalue_correct	freshInt_divLow_pvalue_correct	pastInt_divLow_pvalue_correct	freshInt_divHigh_pvalue_correct	pastInt_divHigh_pvalue_correct
NA	K00003	0	1.0000	53.2414	194.2078	197.5625	248.4660	198.6070	1.0000	196.0480	1.0000	194.9850	1.0000	Gaussian	0.4833	NA	-2.6504	2.9109	0.3633	-3.7757	2.9223	0.1974	4.2762	-3.9392	0.0715	1.2181	-0.3362	-5.4444	3.9499	4.3365	0.9322	0.2104	-4.6378	-1.1353	4.4394	3.8620	0.2971	0.7690	-7.1213	4.9448	3.6536	4.7583	0.0523	0.2996	-2.5899	-2.9112	3.2146	6.9773	0.4211	0.6768	-2.1239	-3.1984	4.1591	4.0434	0.6100	0.4296	-2.0794	-5.8683	3.9499	4.3744	0.5990	0.1809	-7.6452	-0.8076	4.4394	3.8888	0.0862	0.8356	-5.8390	-0.2335	3.6536	4.8091	0.1111	0.9613	-2.9031	-7.8727	3.2300	6.9773	0.3695	0.2602	2.0471	-9.1559	4.1931	4.0434	0.6258	0.0243	0.6955	0.5868	0.9891	0.8941	0.7660	0.7617	0.8427	0.6975	0.9531	0.9680	0.5434	0.6151	0.7640	0.9917	0.8704	0.8572	0.9343	0.8376	0.9057	0.9089	0.8389	0.4256
NA	K00005	0	0.9942	6.2805	34.0300	31.6354	81.7872	31.8760	1.0000	31.7015	1.0000	31.0494	0.9765	Gaussian	0.7267	NA	-0.0439	1.0534	0.9668	-0.5992	1.0576	0.5715	1.4246	-2.7774	-0.1163	1.0979	0.2574	-0.4146	1.4302	1.5698	0.8573	0.7919	-0.4380	0.2641	1.5951	1.3872	0.7838	0.8492	-1.0012	1.5823	1.3291	1.7309	0.4519	0.3615	-0.7583	3.3151	1.1469	2.4882	0.5091	0.1839	-0.5250	0.3984	1.5160	1.4727	0.7294	0.7870	0.1993	-1.5789	1.4302	1.5835	0.8893	0.3197	-3.2757	1.4534	1.5951	1.3968	0.0410	0.2991	-1.2116	0.4565	1.3291	1.7494	0.3628	0.7943	0.2303	-4.4474	1.1524	2.4882	0.8418	0.0750	0.0020	-1.1424	1.5284	1.4727	0.9990	0.4386	0.9869	0.8033	0.9721	0.9798	0.9548	0.7909	0.9550	0.5777	0.9693	0.8429	0.8442	0.7947	0.7941	0.9621	0.8909	0.9715	0.8260	0.7426	0.9361	0.9999	0.9516	0.8435
NA	K00008	0	0.7018	2.2753	15.1554	13.2774	73.5507	13.4150	0.6842	13.5326	0.7442	13.1593	0.6941	Gaussian	0.6964	NA	0.5104	0.7080	0.4716	0.1041	0.7108	0.8836	0.4277	-1.7522	-0.0495	0.2024	0.8551	0.0871	0.9601	1.0538	0.3739	0.9342	1.5990	-0.3087	1.0737	0.9338	0.1376	0.7412	-0.6819	2.5325	0.8869	1.1550	0.4427	0.0292	0.7948	-0.8312	0.7792	1.6904	0.3086	0.6233	1.3629	-0.2887	1.0152	0.9862	0.1806	0.7700	0.8358	-0.7928	0.9601	1.0631	0.3848	0.4565	-0.7072	0.7265	1.0737	0.9402	0.5107	0.4404	-0.1889	0.6042	0.8869	1.1674	0.8315	0.6052	0.5704	-2.0766	0.7829	1.6904	0.4669	0.2204	-0.4609	0.6159	1.0235	0.9862	0.6529	0.5328	0.7542	0.9539	0.8105	0.8125	0.9899	0.8437	0.7734	0.8918	0.9469	0.8817	0.8403	0.9672	0.4652	0.9057	0.8447	0.8762	0.9186	0.8307	0.6914	0.9169	0.9481	0.8772
NA	K00009	0	0.9766	5.2603	31.6988	28.6944	115.4810	27.5855	0.9708	28.3442	1.0000	31.7631	0.9647	Gaussian	0.6133	NA	0.8711	1.4752	0.5553	5.4678	1.4810	0.0003	0.6309	-5.3788	-0.1301	-0.5970	1.0875	0.5791	1.9893	2.1838	0.5851	0.7911	0.9690	0.7975	2.2563	1.9625	0.6679	0.6848	2.6018	-2.0721	1.8531	2.4133	0.1615	0.3913	-0.1795	5.8058	1.6167	3.5081	0.9117	0.0991	-1.1127	2.7219	2.1189	2.0593	0.5999	0.1874	8.1256	2.2125	1.9893	2.2028	0.0001	0.3161	5.3247	5.5778	2.2563	1.9761	0.0190	0.0051	7.3021	2.2959	1.8531	2.4391	0.0001	0.3474	6.0315	2.8755	1.6244	3.5081	0.0002	0.4131	5.2596	5.7046	2.1363	2.0593	0.0144	0.0060	0.7934	0.0552	0.8873	0.0930	0.9546	0.7909	0.9324	0.4766	0.9344	0.4072	0.6656	0.0543	0.8100	0.7860	0.9854	0.1633	0.7698	0.8688	0.9016	0.3844	0.7004	0.3351
NA	K00010	0	0.9883	3.8333	34.7802	31.5682	111.9080	31.6634	0.9825	30.0142	0.9884	33.0006	1.0000	log-Gaussian	0.5907	NA	-0.0595	0.0423	0.1608	0.0093	0.0425	0.8264	0.0200	-0.0238	-0.0019	-0.0070	-0.0395	-0.0844	0.0574	0.0630	0.4921	0.1815	-0.0826	-0.0421	0.0647	0.0563	0.2032	0.4552	-0.0790	-0.0266	0.0535	0.0697	0.1415	0.7031	-0.0735	0.0064	0.0466	0.1012	0.1161	0.9499	-0.0426	-0.0759	0.0608	0.0591	0.4836	0.1999	0.0590	-0.0515	0.0574	0.0635	0.3043	0.4181	-0.0049	0.0203	0.0647	0.0567	0.9394	0.7210	0.0049	0.0169	0.0535	0.0705	0.9272	0.8109	0.0241	-0.0593	0.0469	0.1012	0.6074	0.5584	0.0699	-0.0468	0.0613	0.0591	0.2553	0.4285	0.5545	0.9283	0.8563	0.7870	0.7619	0.8259	0.8127	0.9891	0.8829	0.9417	0.6483	0.9871	0.9367	0.9642	0.7864	0.9127	0.9903	0.9000	0.8604	0.7572	0.7110	0.8379

Number of significant genes

Code

dat_tab <- dat_KOresults %>% 
  mutate(fresh_decision = case_when(freshInt_pvalue_correct < 0.1 ~ "fresh intervention effect significant",
                                    TRUE                           ~ "fresh intervention effect not significant"),
         past_decision  = case_when(pastInt_pvalue_correct < 0.1  ~ "past. intervention effect significant",
                                    TRUE                           ~ "past. intervention effect not significant"))

table(dat_tab$fresh_decision, dat_tab$past_decision) %>% 
  kable() %>% 
  kable_styling()

	past. intervention effect not significant	past. intervention effect significant
fresh intervention effect not significant	2710	75

Significant genes regarding fresh intervention

Code

dat_KOresults %>% 
  filter(freshInt_pvalue_correct < 0.1) %>% 
  arrange(freshInt_pvalue_correct) %>% 
  dplyr::select(-starts_with("pastInt")) %>% 
  kable() %>% 
  kable_styling()

SCFA	gene	relFreq_zeroTPM	relFreq_min10TPM	min_tpmAbd	mean_tpmAbd	median_tpmAbd	max_tpmAbd	baseline_medianTPMAbd	baseline_prevalence	postFresh_medianTPMAbd	postFresh_prevalence	postPast_medianTPMAbd	postPast_prevalence	model	model_deviance	model_meanAbsResid	freshInt_effect	freshInt_se	freshInt_pvalue	phaseEffect	genderEffect	ageEffect	bmiEffect	freshInt_ageLow_effect	freshInt_ageHigh_effect	freshInt_ageLow_se	freshInt_ageHigh_se	freshInt_ageLow_pvalue	freshInt_ageHigh_pvalue	freshInt_sexM_effect	freshInt_sexW_effect	freshInt_sexM_se	freshInt_sexW_se	freshInt_sexM_pvalue	freshInt_sexW_pvalue	freshInt_bmiLow_effect	freshInt_bmiHigh_effect	freshInt_bmiLow_se	freshInt_bmiHigh_se	freshInt_bmiLow_pvalue	freshInt_bmiHigh_pvalue	freshInt_fibLow_effect	freshInt_fibHigh_effect	freshInt_fibLow_se	freshInt_fibHigh_se	freshInt_fibLow_pvalue	freshInt_fibHigh_pvalue	freshInt_divLow_effect	freshInt_divHigh_effect	freshInt_divLow_se	freshInt_divHigh_se	freshInt_divLow_pvalue	freshInt_divHigh_pvalue	freshInt_pvalue_correct	freshInt_ageLow_pvalue_correct	freshInt_ageHigh_pvalue_correct	freshInt_sexM_pvalue_correct	freshInt_sexW_pvalue_correct	freshInt_bmiLow_pvalue_correct	freshInt_bmiHigh_pvalue_correct	freshInt_fibLow_pvalue_correct	freshInt_fibHigh_pvalue_correct	freshInt_divLow_pvalue_correct	freshInt_divHigh_pvalue_correct

Significant genes regarding pasteurized intervention

Code

dat_KOresults %>% 
  filter(pastInt_pvalue_correct < 0.1) %>% 
  arrange(pastInt_pvalue_correct) %>% 
  dplyr::select(-starts_with("freshInt")) %>% 
  kable() %>% 
  kable_styling()

SCFA	gene	relFreq_zeroTPM	relFreq_min10TPM	min_tpmAbd	mean_tpmAbd	median_tpmAbd	max_tpmAbd	baseline_medianTPMAbd	baseline_prevalence	postFresh_medianTPMAbd	postFresh_prevalence	postPast_medianTPMAbd	postPast_prevalence	model	model_deviance	model_meanAbsResid	pastInt_effect	pastInt_se	pastInt_pvalue	phaseEffect	genderEffect	ageEffect	bmiEffect	pastInt_ageLow_effect	pastInt_ageHigh_effect	pastInt_ageLow_se	pastInt_ageHigh_se	pastInt_ageLow_pvalue	pastInt_ageHigh_pvalue	pastInt_sexM_effect	pastInt_sexW_effect	pastInt_sexM_se	pastInt_sexW_se	pastInt_sexM_pvalue	pastInt_sexW_pvalue	pastInt_bmiLow_effect	pastInt_bmiHigh_effect	pastInt_bmiLow_se	pastInt_bmiHigh_se	pastInt_bmiLow_pvalue	pastInt_bmiHigh_pvalue	pastInt_fibLow_effect	pastInt_fibHigh_effect	pastInt_fibLow_se	pastInt_fibHigh_se	pastInt_fibLow_pvalue	pastInt_fibHigh_pvalue	pastInt_divLow_effect	pastInt_divHigh_effect	pastInt_divLow_se	pastInt_divHigh_se	pastInt_divLow_pvalue	pastInt_divHigh_pvalue	pastInt_pvalue_correct	pastInt_ageLow_pvalue_correct	pastInt_ageHigh_pvalue_correct	pastInt_sexM_pvalue_correct	pastInt_sexW_pvalue_correct	pastInt_bmiLow_pvalue_correct	pastInt_bmiHigh_pvalue_correct	pastInt_fibLow_pvalue_correct	pastInt_fibHigh_pvalue_correct	pastInt_divLow_pvalue_correct	pastInt_divHigh_pvalue_correct
NA	K01424	0.0000	1.0000	33.1424	120.3046	121.1000	208.8170	122.1230	1.0000	122.9770	1.0000	117.0730	1.0000	Gaussian	0.5753	NA	-9.2653	2.3022	0.0001	-1.0465	9.1344	0.1137	1.5943	-11.2428	-6.8484	3.1028	3.4360	0.0003	0.0472	-11.5519	-7.5103	3.5006	3.0661	0.0011	0.0149	-10.2908	-7.4894	2.9001	3.8173	0.0005	0.0508	-9.8668	-6.4595	2.5451	5.4970	0.0001	0.2410	-11.7269	-6.9957	3.3224	3.2036	0.0005	0.0298	0.0368	0.2091	0.7130	0.3278	0.4518	0.1038	0.5399	0.1206	0.8372	0.2275	0.4410
NA	K01777	0.0029	0.1988	0.0000	7.8345	7.4021	35.8471	7.5195	0.2456	7.4168	0.1860	6.7787	0.1176	Gaussian	0.7542	NA	-1.3442	0.3130	0.0000	0.0508	0.7071	0.0340	0.3659	-1.1177	-1.6222	0.4232	0.4686	0.0088	0.0006	-1.7717	-1.0165	0.4756	0.4165	0.0002	0.0153	-1.2815	-1.4532	0.3945	0.5193	0.0013	0.0055	-1.4488	-0.8529	0.3457	0.7464	0.0000	0.2542	-1.3704	-1.3134	0.4513	0.4348	0.0026	0.0028	0.0368	0.6399	0.2587	0.2467	0.4544	0.1744	0.3266	0.0888	0.8376	0.3073	0.3073
NA	K01814	0.0000	1.0000	19.9867	82.8668	82.9059	138.9620	84.4132	1.0000	83.4656	1.0000	79.7166	1.0000	Gaussian	0.6235	NA	-5.7533	1.4020	0.0001	0.8560	0.7966	-0.0703	0.6738	-7.5910	-3.5012	1.8907	2.0936	0.0001	0.0956	-5.8871	-5.6507	2.1312	1.8665	0.0061	0.0027	-6.3663	-4.6849	1.7642	2.3222	0.0004	0.0447	-6.8809	-0.4828	1.5412	3.3282	0.0000	0.8848	-8.2949	-3.3911	2.0107	1.9381	0.0000	0.0813	0.0368	0.1078	0.7321	0.4072	0.4004	0.0911	0.5144	0.0440	0.9810	0.1567	0.5507
NA	K02240	0.0029	0.1608	0.0000	5.8848	3.8797	51.7109	4.0799	0.1813	3.7860	0.1628	3.4472	0.1176	log-Gaussian	0.7970	NA	-0.1774	0.0449	0.0001	0.1207	-0.1426	-0.0007	0.0270	-0.1938	-0.1573	0.0608	0.0673	0.0016	0.0202	-0.2426	-0.1274	0.0682	0.0597	0.0004	0.0337	-0.2289	-0.0887	0.0561	0.0738	0.0001	0.2306	-0.1464	-0.3209	0.0491	0.1061	0.0032	0.0027	-0.1525	-0.2014	0.0648	0.0624	0.0193	0.0014	0.0368	0.3383	0.6846	0.2467	0.5466	0.0467	0.7110	0.4390	0.4390	0.4058	0.2889
NA	K02800	0.0000	0.9854	8.3409	34.3059	31.4380	99.9480	29.1504	0.9942	31.3454	0.9651	37.0330	0.9882	Gaussian	0.6669	NA	6.1388	1.4635	0.0000	1.3081	-6.8307	-0.0540	0.0161	9.4735	2.0536	1.9572	2.1672	0.0000	0.3442	5.1728	6.8796	2.2279	1.9511	0.0210	0.0005	7.8027	3.2534	1.8360	2.4167	0.0000	0.1794	6.4926	4.5324	1.6005	3.4562	0.0001	0.1909	6.5718	5.7654	2.1142	2.0376	0.0021	0.0050	0.0368	0.0246	0.8022	0.4836	0.2637	0.0459	0.6782	0.1166	0.8266	0.2895	0.3341
NA	K03483	0.0000	0.8655	1.3906	22.1851	19.6555	84.4915	18.1234	0.8655	19.1580	0.8488	22.5557	0.8824	log-Gaussian	0.6403	NA	0.2027	0.0502	0.0001	0.0329	-0.2000	-0.0062	-0.0182	0.2536	0.1401	0.0677	0.0750	0.0002	0.0628	0.1486	0.2442	0.0763	0.0668	0.0524	0.0003	0.2363	0.1445	0.0631	0.0831	0.0002	0.0832	0.2139	0.1520	0.0549	0.1185	0.0001	0.2010	0.2042	0.2025	0.0726	0.0700	0.0052	0.0041	0.0368	0.1889	0.7172	0.6194	0.2467	0.0717	0.5923	0.1206	0.8285	0.3341	0.3222
NA	K03486	0.0000	0.8070	2.7685	17.5988	15.9824	68.6303	17.1748	0.8246	16.1217	0.8140	14.4540	0.7647	log-Gaussian	0.7372	NA	-0.1517	0.0380	0.0001	-0.0263	-0.1695	0.0005	0.0109	-0.1760	-0.1219	0.0514	0.0569	0.0007	0.0332	-0.1806	-0.1295	0.0579	0.0507	0.0020	0.0111	-0.2099	-0.0510	0.0474	0.0624	0.0000	0.4145	-0.1360	-0.2249	0.0420	0.0906	0.0014	0.0137	-0.1256	-0.1759	0.0550	0.0530	0.0231	0.0010	0.0368	0.2861	0.6943	0.3791	0.4473	0.0459	0.8220	0.3767	0.5956	0.4239	0.2877
NA	K06902	0.0000	1.0000	10.8021	55.6987	54.0527	102.9490	55.3348	1.0000	55.5366	1.0000	52.0884	1.0000	log-Gaussian	0.6504	NA	-0.1210	0.0289	0.0000	0.0384	-0.0360	-0.0034	0.0269	-0.1369	-0.1018	0.0390	0.0431	0.0005	0.0190	-0.1043	-0.1339	0.0440	0.0385	0.0185	0.0006	-0.1445	-0.0805	0.0362	0.0477	0.0001	0.0928	-0.1332	-0.0644	0.0319	0.0689	0.0000	0.3509	-0.1165	-0.1252	0.0418	0.0403	0.0057	0.0021	0.0368	0.2513	0.6846	0.4714	0.2934	0.0533	0.6015	0.0888	0.8533	0.3351	0.2895
NA	K06978	0.0000	0.9152	4.9190	19.0248	17.4046	50.0675	18.4287	0.9240	17.7309	0.9419	15.7752	0.8706	Gaussian	0.7449	NA	-2.3375	0.5937	0.0001	-0.1269	-0.5987	-0.1080	0.3158	-2.4016	-2.2586	0.8021	0.8881	0.0030	0.0116	-1.9224	-2.6559	0.9036	0.7912	0.0343	0.0009	-2.8158	-1.5092	0.7467	0.9828	0.0002	0.1259	-2.5082	-1.5353	0.6553	1.4150	0.0002	0.2789	-3.7213	-1.0495	0.8510	0.8200	0.0000	0.2017	0.0368	0.4183	0.6582	0.5499	0.3224	0.0705	0.6261	0.1206	0.8395	0.0990	0.7117
NA	K10672	0.0000	0.9591	4.7057	26.3704	23.5292	97.7524	24.0854	0.9591	23.2503	0.9535	21.3362	0.9647	log-Gaussian	0.7270	NA	-0.1487	0.0374	0.0001	0.0241	-0.0274	0.0063	0.0311	-0.1761	-0.1151	0.0506	0.0561	0.0006	0.0411	-0.2058	-0.1048	0.0568	0.0498	0.0004	0.0361	-0.2017	-0.0570	0.0466	0.0614	0.0000	0.3539	-0.1460	-0.1611	0.0414	0.0894	0.0005	0.0728	-0.1409	-0.1557	0.0542	0.0522	0.0098	0.0031	0.0368	0.2573	0.7130	0.2467	0.5619	0.0459	0.7899	0.2305	0.7426	0.3755	0.3073
Pyruvate	K13497	0.0000	0.6959	0.4710	14.4480	12.9618	75.9723	11.8396	0.6433	13.0961	0.6512	14.8469	0.8471	log-Gaussian	0.6359	NA	0.1897	0.0447	0.0000	0.0649	0.0114	0.0032	-0.0265	0.2643	0.0982	0.0601	0.0666	0.0000	0.1412	0.1276	0.2374	0.0679	0.0595	0.0613	0.0001	0.2307	0.1189	0.0561	0.0739	0.0001	0.1087	0.1897	0.1905	0.0493	0.1064	0.0001	0.0746	0.1612	0.2165	0.0646	0.0623	0.0132	0.0006	0.0368	0.0353	0.7486	0.6453	0.2064	0.0467	0.6144	0.1206	0.7426	0.3791	0.2275
NA	K16509	0.0058	0.2398	0.0000	8.9212	5.9768	110.6230	6.5589	0.2573	6.2056	0.2558	5.2495	0.1882	log-Gaussian	0.7187	NA	-0.2191	0.0556	0.0001	0.1461	-0.1452	-0.0014	0.0145	-0.2482	-0.1835	0.0752	0.0833	0.0011	0.0285	-0.3239	-0.1388	0.0842	0.0737	0.0002	0.0608	-0.2315	-0.1984	0.0695	0.0914	0.0010	0.0310	-0.1941	-0.3347	0.0610	0.1317	0.0016	0.0116	-0.2331	-0.2074	0.0802	0.0773	0.0040	0.0077	0.0368	0.3059	0.6849	0.2393	0.6432	0.1512	0.4652	0.3947	0.5759	0.3222	0.3548
NA	K17810	0.0000	0.9942	5.2468	35.1354	33.9057	85.0960	35.2136	1.0000	32.9209	0.9884	31.1843	0.9882	log-Gaussian	0.6591	NA	-0.1475	0.0352	0.0000	0.0058	0.0940	-0.0023	0.0202	-0.1739	-0.1153	0.0475	0.0526	0.0003	0.0291	-0.1688	-0.1312	0.0535	0.0469	0.0018	0.0055	-0.1730	-0.1034	0.0442	0.0582	0.0001	0.0769	-0.1506	-0.1332	0.0389	0.0839	0.0001	0.1139	-0.1648	-0.1312	0.0506	0.0488	0.0013	0.0076	0.0368	0.2091	0.6849	0.3776	0.4072	0.0562	0.5816	0.1206	0.7839	0.2889	0.3525
NA	K19157	0.0000	1.0000	60.4201	230.0660	216.4270	893.0340	220.8300	1.0000	219.2595	1.0000	204.2390	1.0000	Gaussian	0.7899	NA	-27.1564	6.5238	0.0000	1.4038	25.1763	-0.0575	-2.6140	-24.5854	-30.3513	8.7949	9.7374	0.0056	0.0020	-21.8393	-31.2335	9.9199	8.6860	0.0286	0.0004	-32.4534	-17.9670	8.2015	10.7950	0.0001	0.0973	-24.8442	-38.1218	7.1507	15.4379	0.0006	0.0142	-18.0207	-35.5162	9.3662	9.0226	0.0555	0.0001	0.0368	0.5799	0.3597	0.5254	0.2467	0.0543	0.6040	0.2659	0.5971	0.5004	0.1567
NA	K19510	0.0000	0.9766	4.7364	32.7387	30.4487	81.2096	31.5631	0.9883	30.7407	0.9651	26.5509	0.9647	log-Gaussian	0.7379	NA	-0.1379	0.0334	0.0000	0.0396	-0.1268	-0.0075	0.0521	-0.1481	-0.1254	0.0452	0.0501	0.0012	0.0129	-0.1671	-0.1155	0.0508	0.0445	0.0011	0.0100	-0.2106	-0.0121	0.0413	0.0544	0.0000	0.8238	-0.1319	-0.1653	0.0369	0.0797	0.0004	0.0390	-0.1339	-0.1416	0.0483	0.0466	0.0060	0.0026	0.0368	0.3059	0.6698	0.3278	0.4353	0.0074	0.9653	0.2105	0.6684	0.3351	0.3073
NA	K19956	0.0000	0.9883	7.3330	34.9770	31.4204	135.0230	29.1937	0.9942	31.1940	0.9884	36.8261	0.9765	log-Gaussian	0.6027	NA	0.1809	0.0455	0.0001	0.0623	-0.1212	-0.0022	-0.0159	0.2738	0.0671	0.0610	0.0676	0.0000	0.3218	0.1796	0.1819	0.0694	0.0608	0.0101	0.0030	0.2114	0.1281	0.0573	0.0754	0.0003	0.0906	0.1852	0.1619	0.0501	0.1082	0.0003	0.1359	0.1852	0.1778	0.0659	0.0635	0.0053	0.0055	0.0368	0.0353	0.7918	0.4362	0.4004	0.0764	0.6015	0.1649	0.7992	0.3341	0.3341
NA	K17202	0.0000	0.2865	0.7368	8.6046	7.3109	32.7252	6.4956	0.2456	7.1767	0.2558	8.4218	0.4000	Gaussian	0.6951	NA	1.7457	0.4463	0.0001	0.0239	-0.0232	0.0071	0.0849	1.8970	1.5602	0.6042	0.6690	0.0019	0.0204	1.0953	2.2444	0.6771	0.5929	0.1070	0.0002	0.9629	3.1014	0.5561	0.7320	0.0846	0.0000	2.3932	-1.2722	0.4823	1.0415	0.0000	0.2230	2.3246	1.2188	0.6409	0.6175	0.0003	0.0495	0.0383	0.3501	0.6846	0.7344	0.2467	0.5949	0.0459	0.0140	0.8307	0.2242	0.4818
NA	K17204	0.0000	0.2281	0.7210	7.7044	6.5511	28.8417	6.0322	0.1930	6.6808	0.2093	7.7010	0.3176	log-Gaussian	0.7032	NA	0.1641	0.0421	0.0001	-0.0031	0.0375	-0.0019	0.0137	0.2070	0.1117	0.0568	0.0629	0.0003	0.0770	0.0890	0.2217	0.0638	0.0558	0.1638	0.0001	0.1105	0.2569	0.0527	0.0693	0.0370	0.0003	0.2134	-0.0654	0.0457	0.0987	0.0000	0.5081	0.2180	0.1153	0.0603	0.0581	0.0004	0.0483	0.0383	0.2091	0.7223	0.7947	0.2064	0.4864	0.0759	0.0273	0.8909	0.2242	0.4779
NA	K01478	0.0000	0.9152	2.8968	22.3548	19.6443	77.0576	20.6079	0.9474	19.8361	0.9070	18.0370	0.8588	log-Gaussian	0.7371	NA	-0.1465	0.0379	0.0001	0.0244	0.0536	0.0049	0.0232	-0.1146	-0.1857	0.0512	0.0566	0.0259	0.0012	-0.1715	-0.1273	0.0577	0.0505	0.0032	0.0123	-0.1926	-0.0668	0.0473	0.0623	0.0001	0.2845	-0.1377	-0.1878	0.0418	0.0903	0.0011	0.0386	-0.0888	-0.1996	0.0545	0.0525	0.1044	0.0002	0.0389	0.6849	0.3059	0.4004	0.4512	0.0467	0.7549	0.3546	0.6684	0.5962	0.1778
NA	K02781	0.0000	0.8977	3.2330	28.8319	25.3120	107.0630	24.4052	0.8947	24.5902	0.8721	28.8944	0.9294	log-Gaussian	0.6018	NA	0.2088	0.0540	0.0001	0.0463	-0.2021	-0.0082	-0.0093	0.2483	0.1605	0.0730	0.0808	0.0008	0.0481	0.1457	0.2573	0.0820	0.0718	0.0766	0.0004	0.2515	0.1345	0.0677	0.0891	0.0002	0.1321	0.2332	0.0963	0.0590	0.1275	0.0001	0.4507	0.1594	0.2557	0.0779	0.0751	0.0418	0.0008	0.0389	0.2861	0.7149	0.6806	0.2467	0.0736	0.6330	0.1206	0.8753	0.4729	0.2584
NA	K02760	0.0000	1.0000	16.5141	79.9727	73.4233	248.7300	77.5212	1.0000	68.4372	1.0000	65.1466	1.0000	log-Gaussian	0.7162	NA	-0.1414	0.0374	0.0002	0.0743	-0.1988	-0.0043	0.0258	-0.1434	-0.1392	0.0507	0.0561	0.0050	0.0137	-0.1765	-0.1145	0.0569	0.0498	0.0021	0.0223	-0.1761	-0.0817	0.0469	0.0617	0.0002	0.1864	-0.1175	-0.2520	0.0409	0.0883	0.0044	0.0046	-0.1361	-0.1463	0.0542	0.0522	0.0126	0.0054	0.0492	0.5609	0.6698	0.3817	0.4855	0.0714	0.6849	0.4513	0.4665	0.3791	0.3341
NA	K07393	0.0029	0.9298	0.0000	25.6790	24.3219	67.5453	24.8859	0.9415	24.8432	0.9302	21.4166	0.9059	Gaussian	0.7107	NA	-3.6654	0.9690	0.0002	0.4776	-2.5730	-0.0216	0.8780	-4.0816	-3.1510	1.3092	1.4496	0.0020	0.0306	-3.9318	-3.4614	1.4758	1.2924	0.0082	0.0079	-4.1480	-2.8371	1.2156	1.6000	0.0007	0.0774	-3.5556	-4.1735	1.0713	2.3133	0.0010	0.0724	-3.0044	-4.2728	1.4007	1.3497	0.0329	0.0017	0.0492	0.3597	0.6849	0.4214	0.4214	0.1299	0.5818	0.3517	0.7426	0.4532	0.2889
NA	K12528	0.0000	0.9766	4.5560	27.8176	26.5937	67.0434	27.9343	0.9825	26.8963	0.9651	23.2623	0.9765	Gaussian	0.7416	NA	-3.0979	0.8231	0.0002	1.5922	-4.5045	-0.1591	0.8118	-3.7212	-2.3350	1.1131	1.2325	0.0009	0.0593	-2.4212	-3.6171	1.2527	1.0970	0.0543	0.0011	-3.5124	-2.3860	1.0312	1.3574	0.0008	0.0799	-3.2771	-2.2655	0.9076	1.9598	0.0004	0.2487	-2.4397	-3.7076	1.1898	1.1465	0.0413	0.0014	0.0500	0.3059	0.7166	0.6261	0.3278	0.1308	0.5857	0.2034	0.8372	0.4717	0.2889
NA	K18692	0.0000	0.1053	0.2272	5.1297	3.5737	47.4198	3.6423	0.1287	3.4486	0.1047	3.3714	0.0588	log-Gaussian	0.7765	NA	-0.1639	0.0437	0.0002	0.1157	-0.1328	-0.0009	0.0203	-0.1753	-0.1498	0.0592	0.0655	0.0033	0.0230	-0.2708	-0.0819	0.0659	0.0577	0.0001	0.1569	-0.1966	-0.1075	0.0546	0.0719	0.0004	0.1359	-0.1310	-0.3162	0.0477	0.1030	0.0065	0.0024	-0.1458	-0.1814	0.0631	0.0608	0.0216	0.0031	0.0505	0.4366	0.6849	0.2064	0.7893	0.0923	0.6349	0.5135	0.4339	0.4189	0.3073
NA	K02245	0.0117	0.1228	0.0000	5.3248	3.5439	72.9346	3.7430	0.1404	3.3975	0.1279	3.2111	0.0824	log-Gaussian	0.7796	NA	-0.1757	0.0471	0.0002	0.1096	-0.1423	-0.0010	0.0250	-0.2065	-0.1379	0.0637	0.0705	0.0013	0.0514	-0.2548	-0.1151	0.0714	0.0625	0.0004	0.0667	-0.2320	-0.0786	0.0588	0.0774	0.0001	0.3109	-0.1429	-0.3274	0.0514	0.1109	0.0058	0.0035	-0.1452	-0.2045	0.0681	0.0656	0.0338	0.0020	0.0519	0.3101	0.7149	0.2467	0.6559	0.0543	0.7678	0.5020	0.4434	0.4532	0.2895
NA	K00009	0.0000	0.9766	5.2603	31.6988	28.6944	115.4810	27.5855	0.9708	28.3442	1.0000	31.7631	0.9647	Gaussian	0.6133	NA	5.4678	1.4810	0.0003	0.6309	-5.3788	-0.1301	-0.5970	8.1256	2.2125	1.9893	2.2028	0.0001	0.3161	5.3247	5.5778	2.2563	1.9761	0.0190	0.0051	7.3021	2.2959	1.8531	2.4391	0.0001	0.3474	6.0315	2.8755	1.6244	3.5081	0.0002	0.4131	5.2596	5.7046	2.1363	2.0593	0.0144	0.0060	0.0552	0.0930	0.7909	0.4766	0.4072	0.0543	0.7860	0.1633	0.8688	0.3844	0.3351
NA	K01042	0.0000	0.9064	2.1992	20.2673	18.8658	67.5662	19.0928	0.9357	19.7120	0.8953	16.4124	0.8588	log-Gaussian	0.7483	NA	-0.1266	0.0343	0.0003	0.0268	-0.0267	0.0038	0.0331	-0.0906	-0.1706	0.0463	0.0512	0.0514	0.0010	-0.1553	-0.1046	0.0522	0.0457	0.0032	0.0229	-0.1791	-0.0357	0.0427	0.0562	0.0000	0.5254	-0.1211	-0.1518	0.0379	0.0818	0.0016	0.0646	-0.0840	-0.1657	0.0494	0.0476	0.0900	0.0006	0.0552	0.7149	0.3059	0.4004	0.4912	0.0460	0.8794	0.3947	0.7413	0.5731	0.2275
NA	K01497	0.0000	0.3889	1.1883	9.5502	8.6160	34.5050	8.0224	0.3392	8.8291	0.4070	9.5833	0.4706	log-Gaussian	0.6102	NA	0.1697	0.0461	0.0003	0.0357	-0.0903	-0.0058	0.0106	0.2304	0.0952	0.0620	0.0687	0.0002	0.1671	0.1370	0.1948	0.0701	0.0614	0.0518	0.0017	0.1806	0.1506	0.0580	0.0764	0.0021	0.0497	0.1949	0.0528	0.0507	0.1095	0.0002	0.6302	0.1599	0.1794	0.0666	0.0642	0.0171	0.0056	0.0552	0.1982	0.7544	0.6165	0.3697	0.2051	0.5332	0.1206	0.9197	0.3965	0.3347
NA	K19509	0.0000	0.8626	1.4455	21.5168	18.5855	79.5557	19.7792	0.8655	18.9192	0.8837	16.5391	0.8353	log-Gaussian	0.8189	NA	-0.1369	0.0372	0.0003	0.1039	-0.3228	-0.0088	0.0332	-0.1814	-0.0825	0.0502	0.0555	0.0004	0.1388	-0.1589	-0.1200	0.0566	0.0496	0.0054	0.0163	-0.2049	-0.0191	0.0464	0.0611	0.0000	0.7545	-0.1093	-0.2655	0.0410	0.0885	0.0081	0.0029	-0.1276	-0.1455	0.0539	0.0519	0.0186	0.0054	0.0552	0.2091	0.7486	0.4072	0.4584	0.0459	0.9549	0.5397	0.4390	0.4025	0.3341
NA	K01299	0.0000	0.9766	4.2924	30.4604	28.8591	61.5233	29.9912	0.9883	28.8573	0.9651	26.7801	0.9647	log-Gaussian	0.6838	NA	-0.1172	0.0321	0.0003	0.0208	0.0842	0.0031	0.0316	-0.1374	-0.0928	0.0433	0.0479	0.0017	0.0540	-0.1622	-0.0828	0.0488	0.0427	0.0010	0.0539	-0.1571	-0.0481	0.0403	0.0530	0.0001	0.3654	-0.1031	-0.1835	0.0354	0.0765	0.0039	0.0172	-0.1200	-0.1140	0.0459	0.0443	0.0095	0.0105	0.0573	0.3383	0.7149	0.3224	0.6261	0.0562	0.7970	0.4434	0.6278	0.3718	0.3757
NA	K01914	0.0000	1.0000	36.6096	108.3381	109.3120	164.6550	110.9380	1.0000	108.7070	1.0000	103.5820	1.0000	Gaussian	0.5573	NA	-5.5959	1.5386	0.0003	0.5691	-0.4816	0.1323	1.1169	-7.0257	-3.8445	2.0788	2.3021	0.0008	0.0961	-5.3836	-5.7574	2.3284	2.0395	0.0215	0.0051	-7.9766	-1.4786	1.9190	2.5259	0.0000	0.5588	-5.8432	-4.4363	1.7017	3.6755	0.0007	0.2285	-5.9425	-5.2942	2.2267	2.1470	0.0081	0.0143	0.0573	0.2893	0.7321	0.4852	0.4072	0.0467	0.8886	0.2736	0.8315	0.3610	0.3832
NA	K06890	0.0000	0.9942	7.2313	39.4151	37.2706	99.5786	39.6014	0.9942	34.7195	1.0000	36.4245	0.9882	log-Gaussian	0.7294	NA	-0.1238	0.0340	0.0003	0.0134	0.0056	0.0008	0.0182	-0.1759	-0.0602	0.0456	0.0505	0.0001	0.2344	-0.1281	-0.1205	0.0516	0.0452	0.0137	0.0081	-0.1265	-0.1193	0.0428	0.0563	0.0034	0.0350	-0.1044	-0.2138	0.0374	0.0808	0.0057	0.0086	-0.1915	-0.0614	0.0487	0.0470	0.0001	0.1919	0.0573	0.1348	0.7667	0.4518	0.4214	0.2728	0.4819	0.5001	0.5397	0.1567	0.7041
NA	K00030	0.0000	0.9152	1.0457	21.1786	20.1527	56.5064	20.9401	0.9415	19.3735	0.9070	18.7931	0.8706	log-Gaussian	0.6722	NA	-0.1373	0.0378	0.0003	0.0380	-0.0419	-0.0018	0.0306	-0.1367	-0.1382	0.0512	0.0567	0.0081	0.0155	-0.1469	-0.1299	0.0576	0.0504	0.0113	0.0105	-0.1685	-0.0833	0.0475	0.0626	0.0005	0.1843	-0.1604	-0.0292	0.0417	0.0900	0.0001	0.7462	-0.1713	-0.1050	0.0544	0.0524	0.0018	0.0462	0.0577	0.6399	0.6698	0.4476	0.4425	0.1038	0.6828	0.1206	0.9482	0.2889	0.4779
NA	K06971	0.0029	0.3655	0.0000	9.8625	8.3033	33.2740	9.0905	0.4035	8.1373	0.3488	7.4715	0.3059	log-Gaussian	0.6705	NA	-0.1488	0.0411	0.0004	-0.0010	0.0088	-0.0007	0.0209	-0.2277	-0.0523	0.0552	0.0611	0.0001	0.3929	-0.1784	-0.1262	0.0626	0.0548	0.0047	0.0221	-0.1869	-0.0827	0.0516	0.0679	0.0003	0.2243	-0.1491	-0.1471	0.0453	0.0978	0.0011	0.1337	-0.1238	-0.1712	0.0593	0.0572	0.0378	0.0030	0.0583	0.0930	0.8160	0.4072	0.4852	0.0897	0.7085	0.3546	0.7992	0.4619	0.3073
NA	K23121	0.0000	0.2544	0.9395	8.4897	7.7824	25.2666	7.9853	0.2865	7.8147	0.2674	7.2420	0.1765	log-Gaussian	0.5815	NA	-0.1323	0.0367	0.0004	-0.0271	0.0268	-0.0008	0.0243	-0.1345	-0.1294	0.0493	0.0546	0.0069	0.0186	-0.1535	-0.1160	0.0558	0.0489	0.0064	0.0184	-0.1783	-0.0524	0.0459	0.0604	0.0001	0.3868	-0.1331	-0.1278	0.0405	0.0874	0.0011	0.1451	-0.1612	-0.1052	0.0530	0.0511	0.0026	0.0405	0.0587	0.6063	0.6846	0.4072	0.4710	0.0577	0.8100	0.3546	0.8047	0.3073	0.4709
NA	K02745	0.0029	0.4006	0.0000	9.7838	8.2668	53.0323	8.7820	0.4327	7.7779	0.3953	7.4664	0.3412	log-Gaussian	0.7518	NA	-0.1820	0.0512	0.0005	0.0749	-0.1737	0.0026	0.0638	-0.2128	-0.1445	0.0692	0.0766	0.0023	0.0604	-0.2391	-0.1383	0.0779	0.0682	0.0024	0.0437	-0.1850	-0.1772	0.0645	0.0849	0.0045	0.0380	-0.1701	-0.2377	0.0566	0.1223	0.0029	0.0529	-0.2066	-0.1598	0.0741	0.0714	0.0057	0.0261	0.0698	0.3815	0.7172	0.3993	0.5861	0.3228	0.4906	0.4390	0.7119	0.3351	0.4288
NA	K00020	0.0000	0.9854	4.0023	25.2860	23.8846	71.0569	24.9625	1.0000	22.6221	0.9767	23.1333	0.9647	log-Gaussian	0.6701	NA	-0.1109	0.0317	0.0005	0.0157	-0.0858	0.0021	0.0312	-0.0817	-0.1466	0.0428	0.0474	0.0575	0.0022	-0.0996	-0.1195	0.0483	0.0423	0.0400	0.0051	-0.1495	-0.0439	0.0397	0.0522	0.0002	0.4010	-0.1150	-0.0908	0.0346	0.0748	0.0010	0.2257	-0.0680	-0.1501	0.0456	0.0439	0.1366	0.0007	0.0805	0.7149	0.3778	0.5716	0.4072	0.0705	0.8158	0.3517	0.8312	0.6365	0.2538
NA	K08177	0.0000	1.0000	12.1593	41.1144	39.9032	85.0838	41.9585	1.0000	40.3902	1.0000	38.2433	1.0000	Gaussian	0.6433	NA	-3.7143	1.0615	0.0005	1.3324	1.1857	-0.1371	0.5736	-3.0444	-4.5395	1.4339	1.5878	0.0347	0.0046	-5.1183	-2.6375	1.6114	1.4112	0.0017	0.0627	-4.6287	-2.1343	1.3326	1.7541	0.0006	0.2248	-3.0249	-6.9340	1.1698	2.5261	0.0102	0.0065	-5.4636	-2.0742	1.5264	1.4711	0.0004	0.1597	0.0805	0.7021	0.5460	0.3697	0.6485	0.1152	0.7085	0.5719	0.5135	0.2275	0.6668
NA	K06180	0.0000	1.0000	64.9669	215.6466	215.9980	274.0710	218.0880	1.0000	218.3885	1.0000	212.1470	1.0000	Gaussian	0.4837	NA	-9.0821	2.6094	0.0006	3.0099	2.1668	0.3503	1.4619	-8.6135	-9.6698	3.5152	3.8931	0.0149	0.0136	-15.2439	-4.3540	3.9419	3.4531	0.0001	0.2084	-9.7075	-8.0176	3.2775	4.3142	0.0033	0.0642	-8.3505	-12.5229	2.8816	6.2253	0.0041	0.0452	-4.3305	-13.4981	3.7424	3.6095	0.2482	0.0002	0.0814	0.6698	0.6698	0.2393	0.8171	0.2720	0.5646	0.4434	0.6851	0.7556	0.1778
NA	K07029	0.0000	1.0000	67.6451	239.9732	239.9145	327.9390	240.3360	1.0000	241.4220	1.0000	238.9860	1.0000	Gaussian	0.5379	NA	-11.2976	3.2537	0.0006	1.8469	-0.2456	0.1686	2.1968	-10.1771	-12.6772	4.3936	4.8657	0.0213	0.0097	-15.4679	-8.0966	4.9293	4.3176	0.0019	0.0618	-13.7588	-7.0574	4.0774	5.3670	0.0008	0.1896	-10.2523	-16.1546	3.5918	7.7583	0.0046	0.0383	-6.5031	-15.7759	4.6891	4.5211	0.1666	0.0006	0.0814	0.6846	0.6399	0.3776	0.6455	0.1410	0.6878	0.4665	0.6684	0.6755	0.2275
NA	K10794	0.0000	0.1871	0.9772	7.5465	6.8353	30.1187	7.0050	0.2047	6.9130	0.2093	6.1879	0.1294	Gaussian	0.6919	NA	-1.1920	0.3426	0.0006	0.0144	0.3579	0.0454	0.3652	-1.1338	-1.2639	0.4637	0.5135	0.0151	0.0145	-1.8034	-0.7232	0.5196	0.4550	0.0006	0.1132	-1.1991	-1.1798	0.4319	0.5685	0.0059	0.0389	-1.2147	-1.0826	0.3787	0.8178	0.0015	0.1867	-1.3495	-1.0404	0.4945	0.4765	0.0068	0.0299	0.0814	0.6698	0.6698	0.2934	0.7452	0.3325	0.4937	0.3947	0.8266	0.3516	0.4410
NA	K01281	0.0000	0.2573	1.4890	8.5402	7.2181	61.0040	7.5921	0.2865	7.2353	0.2442	6.0575	0.2118	log-Gaussian	0.6935	NA	-0.1349	0.0391	0.0007	0.0194	0.0187	0.0050	0.0182	-0.1731	-0.0881	0.0528	0.0585	0.0012	0.1331	-0.1831	-0.0980	0.0594	0.0520	0.0023	0.0607	-0.1623	-0.0879	0.0488	0.0643	0.0010	0.1729	-0.1195	-0.2068	0.0432	0.0933	0.0060	0.0275	-0.1245	-0.1449	0.0566	0.0545	0.0286	0.0084	0.0834	0.3059	0.7486	0.3946	0.6432	0.1526	0.6736	0.5027	0.6469	0.4366	0.3662
NA	K07251	0.0000	0.3626	0.9618	9.5772	7.9222	63.5507	8.3308	0.3918	8.2335	0.3721	6.6854	0.2941	log-Gaussian	0.7692	NA	-0.1494	0.0433	0.0007	0.0573	-0.0535	-0.0088	0.0276	-0.1221	-0.1831	0.0585	0.0647	0.0378	0.0050	-0.1197	-0.1722	0.0660	0.0578	0.0707	0.0032	-0.2212	-0.0251	0.0539	0.0710	0.0001	0.7238	-0.1145	-0.3119	0.0476	0.1028	0.0169	0.0026	-0.1556	-0.1435	0.0627	0.0604	0.0137	0.0182	0.0834	0.7064	0.5609	0.6667	0.4004	0.0467	0.9444	0.6278	0.4390	0.3800	0.4017
NA	K07722	0.0000	0.6023	1.5045	12.5528	11.1472	64.9527	11.8575	0.6257	11.8009	0.6512	10.1834	0.5059	log-Gaussian	0.7315	NA	-0.1416	0.0410	0.0006	0.0174	0.0456	0.0083	0.0133	-0.1223	-0.1654	0.0554	0.0614	0.0283	0.0075	-0.1412	-0.1419	0.0624	0.0547	0.0246	0.0100	-0.1962	-0.0471	0.0513	0.0675	0.0002	0.4861	-0.1425	-0.1378	0.0453	0.0978	0.0019	0.1603	-0.1211	-0.1603	0.0593	0.0571	0.0421	0.0054	0.0834	0.6849	0.6209	0.5089	0.4353	0.0638	0.8607	0.4063	0.8200	0.4745	0.3341
NA	K01258	0.0000	1.0000	38.2694	159.8186	160.0600	245.8310	161.6090	1.0000	160.6395	1.0000	154.2200	1.0000	Gaussian	0.5547	NA	-8.0070	2.3449	0.0007	3.5741	-6.7356	0.0962	1.5112	-8.2606	-7.7011	3.1748	3.5159	0.0098	0.0293	-8.8966	-7.3258	3.5717	3.1285	0.0133	0.0199	-10.2960	-4.0556	2.9457	3.8773	0.0006	0.2965	-7.6179	-9.7691	2.5753	5.5626	0.0034	0.0802	-7.6192	-8.3807	3.3934	3.2718	0.0255	0.0110	0.0836	0.6399	0.6849	0.4518	0.4832	0.1119	0.7613	0.4434	0.7497	0.4275	0.3779
NA	K01259	0.0029	0.4035	0.0000	11.2897	7.9165	57.1981	7.8159	0.4152	9.1865	0.4419	7.2522	0.3412	log-Gaussian	0.8378	NA	-0.1417	0.0414	0.0007	0.0153	-0.2142	0.0047	0.0322	-0.1202	-0.1679	0.0561	0.0621	0.0330	0.0073	-0.1541	-0.1322	0.0631	0.0553	0.0153	0.0175	-0.1489	-0.1291	0.0522	0.0688	0.0047	0.0615	-0.1273	-0.2078	0.0456	0.0984	0.0056	0.0356	-0.1154	-0.1669	0.0598	0.0576	0.0549	0.0041	0.0836	0.6943	0.6158	0.4544	0.4666	0.3228	0.5634	0.5001	0.6684	0.4984	0.3222
NA	K04651	0.0000	1.0000	18.7232	76.5684	71.9150	222.9510	74.1096	1.0000	71.9150	1.0000	67.3661	1.0000	log-Gaussian	0.7074	NA	-0.1062	0.0310	0.0007	0.0187	0.1403	0.0076	0.0241	-0.0844	-0.1331	0.0419	0.0464	0.0452	0.0045	-0.1500	-0.0727	0.0471	0.0413	0.0016	0.0795	-0.1265	-0.0712	0.0391	0.0514	0.0014	0.1676	-0.1004	-0.1335	0.0343	0.0741	0.0037	0.0728	-0.0947	-0.1169	0.0447	0.0431	0.0350	0.0071	0.0836	0.7130	0.5460	0.3697	0.6864	0.1753	0.6696	0.4434	0.7426	0.4566	0.3525
NA	K09963	0.0000	0.4240	0.6694	10.6018	8.8961	47.1530	9.0755	0.4386	9.2760	0.4651	8.1133	0.3529	log-Gaussian	0.7256	NA	-0.1476	0.0429	0.0007	0.0565	-0.1545	-0.0050	0.0208	-0.1724	-0.1175	0.0580	0.0642	0.0032	0.0682	-0.2116	-0.0986	0.0652	0.0571	0.0013	0.0852	-0.2007	-0.0559	0.0537	0.0707	0.0002	0.4305	-0.1152	-0.2978	0.0468	0.1010	0.0144	0.0035	-0.1520	-0.1440	0.0621	0.0598	0.0151	0.0169	0.0836	0.4321	0.7222	0.3485	0.6965	0.0717	0.8334	0.6017	0.4434	0.3886	0.3945
NA	K19267	0.0029	0.3684	0.0000	9.5612	7.8405	47.0339	8.3720	0.3860	7.2988	0.3488	6.3760	0.3529	log-Gaussian	0.7526	NA	-0.1586	0.0462	0.0007	0.0947	0.0104	-0.0074	0.0229	-0.0988	-0.2319	0.0624	0.0690	0.1145	0.0009	-0.1280	-0.1821	0.0704	0.0617	0.0703	0.0034	-0.2470	-0.0059	0.0570	0.0751	0.0000	0.9369	-0.1114	-0.3783	0.0506	0.1092	0.0285	0.0006	-0.1219	-0.1927	0.0669	0.0644	0.0694	0.0030	0.0836	0.7449	0.3047	0.6644	0.4004	0.0459	0.9885	0.6474	0.2659	0.5283	0.3073
NA	K05346	0.0000	0.9561	1.2041	32.0852	29.0099	95.7052	27.7434	0.9532	26.5915	0.9419	32.9738	0.9765	log-Gaussian	0.6799	NA	0.1634	0.0481	0.0008	0.0186	-0.1152	-0.0080	-0.0029	0.2148	0.1003	0.0649	0.0719	0.0011	0.1639	0.1318	0.1876	0.0733	0.0642	0.0732	0.0038	0.1607	0.1678	0.0606	0.0797	0.0085	0.0362	0.2119	-0.0618	0.0526	0.1136	0.0001	0.5865	0.1697	0.1590	0.0693	0.0668	0.0150	0.0180	0.0849	0.3059	0.7530	0.6759	0.4061	0.3722	0.4844	0.1166	0.9069	0.3886	0.4017
NA	K10796	0.0029	0.1140	0.0000	6.3026	5.9366	21.5495	6.0836	0.1228	6.0554	0.1512	5.2583	0.0588	log-Gaussian	0.6475	NA	-0.1362	0.0401	0.0008	-0.0214	-0.0017	0.0055	0.0337	-0.1204	-0.1554	0.0542	0.0601	0.0273	0.0102	-0.1983	-0.0885	0.0608	0.0533	0.0013	0.0979	-0.1772	-0.0652	0.0503	0.0662	0.0005	0.3259	-0.1217	-0.2034	0.0443	0.0957	0.0064	0.0344	-0.0549	-0.2111	0.0574	0.0553	0.3399	0.0002	0.0849	0.6849	0.6399	0.3433	0.7217	0.1076	0.7751	0.5135	0.6599	0.7973	0.1778
NA	K18890	0.0000	0.9678	0.6334	25.4065	23.4384	58.9354	24.7482	0.9883	23.0303	0.9651	22.8075	0.9294	log-Gaussian	0.7226	NA	-0.1203	0.0353	0.0008	0.0348	0.1346	-0.0063	0.0296	-0.1268	-0.1122	0.0478	0.0530	0.0085	0.0350	-0.1341	-0.1097	0.0538	0.0471	0.0133	0.0207	-0.1460	-0.0758	0.0444	0.0584	0.0011	0.1952	-0.1135	-0.1526	0.0390	0.0841	0.0039	0.0707	-0.1939	-0.0518	0.0507	0.0488	0.0002	0.2897	0.0849	0.6399	0.7034	0.4518	0.4836	0.1620	0.6919	0.4434	0.7426	0.1778	0.7764
NA	K02082	0.0000	0.4708	0.4768	10.9479	9.3409	45.3290	9.6755	0.4912	9.4341	0.4884	8.5374	0.4118	log-Gaussian	0.7714	NA	-0.1594	0.0472	0.0008	0.0475	-0.0552	-0.0007	0.0583	-0.1555	-0.1645	0.0637	0.0706	0.0153	0.0205	-0.1840	-0.1406	0.0718	0.0629	0.0110	0.0262	-0.1850	-0.1152	0.0594	0.0782	0.0020	0.1418	-0.1287	-0.3028	0.0519	0.1122	0.0139	0.0074	-0.1505	-0.1677	0.0683	0.0658	0.0283	0.0113	0.0860	0.6698	0.6846	0.4473	0.5210	0.2051	0.6483	0.5956	0.5283	0.4350	0.3791
NA	K02769	0.0029	0.7632	0.0000	20.7022	18.0266	81.5632	16.3253	0.7661	18.1801	0.7442	22.1653	0.7765	log-Gaussian	0.7197	NA	0.1921	0.0568	0.0008	0.1095	-0.1216	-0.0073	-0.0183	0.2389	0.1343	0.0765	0.0847	0.0020	0.1140	0.1526	0.2223	0.0864	0.0756	0.0785	0.0036	0.2227	0.1390	0.0715	0.0941	0.0021	0.1409	0.1903	0.2014	0.0626	0.1351	0.0026	0.1373	0.1928	0.1926	0.0822	0.0792	0.0197	0.0156	0.0860	0.3597	0.7433	0.6843	0.4055	0.2051	0.6462	0.4390	0.7992	0.4091	0.3886
NA	K00068	0.0000	0.8626	4.3246	19.4311	17.5620	64.9188	16.5446	0.8363	17.5824	0.8721	20.1357	0.9059	log-Gaussian	0.6073	NA	0.1466	0.0437	0.0009	0.0449	-0.0876	-0.0012	-0.0070	0.2305	0.0437	0.0586	0.0649	0.0001	0.5014	0.1088	0.1755	0.0664	0.0581	0.1023	0.0028	0.1582	0.1262	0.0550	0.0724	0.0044	0.0826	0.1799	-0.0075	0.0477	0.1029	0.0002	0.9422	0.1658	0.1300	0.0631	0.0609	0.0091	0.0336	0.0888	0.1195	0.8581	0.7252	0.4004	0.3202	0.5904	0.1377	0.9890	0.3704	0.4532
NA	K03833	0.0000	0.8830	2.4043	18.4745	17.1400	52.2558	17.5496	0.9123	17.7206	0.8721	16.2770	0.8353	log-Gaussian	0.7519	NA	-0.1085	0.0322	0.0009	0.0223	-0.0484	0.0028	0.0345	-0.0730	-0.1519	0.0435	0.0482	0.0945	0.0018	-0.1332	-0.0895	0.0491	0.0430	0.0071	0.0382	-0.1507	-0.0355	0.0403	0.0530	0.0002	0.5030	-0.1132	-0.0861	0.0356	0.0769	0.0017	0.2641	-0.0678	-0.1459	0.0464	0.0447	0.1452	0.0013	0.0888	0.7321	0.3414	0.4171	0.5716	0.0717	0.8708	0.3947	0.8395	0.6470	0.2889
NA	K21577	0.0000	0.1053	1.2826	6.4421	5.9219	20.3011	6.1947	0.1228	6.0014	0.1163	5.1688	0.0588	log-Gaussian	0.7194	NA	-0.1095	0.0326	0.0009	-0.0225	-0.0160	0.0038	0.0274	-0.0954	-0.1266	0.0441	0.0488	0.0314	0.0101	-0.1334	-0.0912	0.0496	0.0435	0.0077	0.0368	-0.1566	-0.0280	0.0408	0.0537	0.0002	0.6020	-0.1179	-0.0702	0.0361	0.0779	0.0012	0.3682	-0.1095	-0.1093	0.0472	0.0455	0.0211	0.0170	0.0888	0.6882	0.6399	0.4214	0.5643	0.0638	0.9044	0.3631	0.8572	0.4189	0.3955
NA	K01304	0.0000	0.9942	8.6652	45.6939	44.1047	111.1400	45.1066	1.0000	45.7141	0.9884	39.4443	0.9882	log-Gaussian	0.5747	NA	-0.1146	0.0342	0.0009	0.0392	0.0517	-0.0005	0.0312	-0.1461	-0.0762	0.0460	0.0509	0.0016	0.1357	-0.1403	-0.0949	0.0521	0.0456	0.0076	0.0387	-0.1260	-0.0950	0.0430	0.0566	0.0036	0.0944	-0.1132	-0.1214	0.0378	0.0816	0.0030	0.1382	-0.1151	-0.1142	0.0495	0.0477	0.0208	0.0174	0.0897	0.3383	0.7486	0.4186	0.5716	0.2862	0.6040	0.4390	0.7994	0.4172	0.3999
NA	K01239	0.0000	1.0000	21.7262	91.8180	87.4888	191.2760	91.2652	1.0000	86.6266	1.0000	82.8117	1.0000	log-Gaussian	0.5037	NA	-0.0965	0.0289	0.0010	0.0147	0.0243	-0.0015	0.0142	-0.0903	-0.1042	0.0390	0.0432	0.0215	0.0166	-0.0948	-0.0977	0.0440	0.0386	0.0322	0.0118	-0.1071	-0.0783	0.0363	0.0478	0.0035	0.1029	-0.0903	-0.1254	0.0320	0.0691	0.0051	0.0704	-0.0594	-0.1309	0.0417	0.0402	0.1555	0.0013	0.0898	0.6846	0.6698	0.5355	0.4476	0.2769	0.6093	0.4715	0.7426	0.6607	0.2889
NA	K02526	0.0000	0.6579	2.4754	14.9242	12.7851	52.0248	13.3288	0.6842	12.6246	0.6047	12.1132	0.6588	log-Gaussian	0.7785	NA	-0.1294	0.0387	0.0010	0.0116	-0.0445	-0.0023	0.0533	-0.1152	-0.1470	0.0523	0.0579	0.0284	0.0117	-0.1131	-0.1419	0.0590	0.0516	0.0561	0.0064	-0.1472	-0.0987	0.0487	0.0641	0.0028	0.1250	-0.1217	-0.1654	0.0428	0.0924	0.0048	0.0747	-0.0902	-0.1654	0.0559	0.0539	0.1077	0.0024	0.0898	0.6849	0.6582	0.6324	0.4072	0.2353	0.6253	0.4715	0.7426	0.5996	0.2988
NA	K01079	0.0000	0.8918	0.8281	24.4933	21.5427	81.1856	23.2068	0.9064	22.1598	0.9070	19.5643	0.8471	log-Gaussian	0.6907	NA	-0.1527	0.0460	0.0010	-0.0085	0.0568	-0.0018	0.0390	-0.1856	-0.1126	0.0620	0.0686	0.0030	0.1020	-0.1392	-0.1631	0.0700	0.0613	0.0479	0.0083	-0.1850	-0.0970	0.0577	0.0760	0.0015	0.2032	-0.1425	-0.2011	0.0507	0.1094	0.0053	0.0672	-0.2042	-0.1061	0.0659	0.0635	0.0021	0.0958	0.0903	0.4183	0.7322	0.5978	0.4214	0.1845	0.6955	0.4838	0.7426	0.2895	0.5836
NA	K01142	0.0000	1.0000	42.0330	152.8160	153.3920	205.2690	154.6360	1.0000	154.6825	1.0000	149.4610	1.0000	Gaussian	0.5047	NA	-6.9164	2.0826	0.0010	1.7029	2.1126	0.3183	1.0194	-7.7870	-5.8544	2.8149	3.1174	0.0060	0.0614	-10.4908	-4.1731	3.1570	2.7655	0.0010	0.1324	-6.4255	-7.7733	2.6230	3.4526	0.0149	0.0251	-6.6104	-8.3496	2.3032	4.9755	0.0044	0.0944	-6.3297	-7.4533	3.0095	2.9023	0.0363	0.0108	0.0903	0.5964	0.7172	0.3224	0.7705	0.4063	0.4553	0.4517	0.7625	0.4567	0.3757
NA	K10563	0.0000	0.6228	2.0462	14.1461	12.0160	63.7619	12.6007	0.6608	11.8714	0.6279	10.6932	0.5412	log-Gaussian	0.7686	NA	-0.1329	0.0400	0.0010	0.0927	-0.1134	0.0061	0.0281	-0.0928	-0.1817	0.0537	0.0595	0.0853	0.0025	-0.2158	-0.0694	0.0604	0.0529	0.0004	0.1910	-0.1552	-0.0948	0.0498	0.0655	0.0020	0.1492	-0.1194	-0.1949	0.0441	0.0952	0.0072	0.0415	-0.1414	-0.1257	0.0578	0.0557	0.0151	0.0250	0.0903	0.7239	0.3982	0.2467	0.8093	0.2051	0.6548	0.5283	0.6751	0.3886	0.4271
NA	K01200	0.0000	1.0000	17.4383	71.9868	68.7343	210.7560	69.7969	1.0000	67.2816	1.0000	69.5399	1.0000	log-Gaussian	0.6383	NA	-0.1188	0.0363	0.0012	0.0218	-0.0321	-0.0046	0.0071	-0.1883	-0.0340	0.0483	0.0535	0.0001	0.5256	-0.1378	-0.1043	0.0553	0.0484	0.0133	0.0320	-0.1275	-0.1039	0.0457	0.0601	0.0056	0.0851	-0.1158	-0.1336	0.0401	0.0866	0.0042	0.1240	-0.1506	-0.0894	0.0524	0.0505	0.0044	0.0782	0.0953	0.1248	0.8687	0.4518	0.5337	0.3266	0.5949	0.4454	0.7931	0.3334	0.5467
NA	K01620	0.0000	1.0000	37.7395	134.9037	135.8880	187.6610	136.8110	1.0000	135.5140	1.0000	134.7930	1.0000	Gaussian	0.5675	NA	-5.8977	1.7999	0.0012	2.5086	-4.1186	-0.1261	1.3453	-6.3927	-5.2951	2.4351	2.6966	0.0091	0.0506	-5.0639	-6.5373	2.7377	2.3980	0.0654	0.0068	-7.8734	-2.4800	2.2575	2.9715	0.0006	0.4047	-5.6264	-7.1596	1.9854	4.2884	0.0049	0.0962	-6.1329	-5.6562	2.6033	2.5100	0.0192	0.0250	0.0953	0.6399	0.7149	0.6559	0.4171	0.1130	0.8182	0.4715	0.7633	0.4056	0.4271
NA	K02759	0.0000	0.9971	9.1115	55.6837	48.9851	188.0850	52.1957	1.0000	46.6576	1.0000	45.5281	0.9882	Gaussian	0.7483	NA	-6.9191	2.1199	0.0012	4.2451	-7.9718	-0.1818	2.4292	-6.9749	-6.8705	2.8539	3.1599	0.0152	0.0306	-7.8244	-6.2256	3.2292	2.8277	0.0161	0.0286	-7.5266	-5.8777	2.6646	3.5072	0.0051	0.0950	-6.7349	-7.7195	2.3273	5.0249	0.0041	0.1257	-8.7032	-5.2276	3.0555	2.9439	0.0047	0.0769	0.0953	0.6698	0.6849	0.4557	0.5254	0.3266	0.6040	0.4454	0.7931	0.3341	0.5467
NA	K02773	0.0000	0.9240	2.1438	31.4024	27.3372	115.6650	25.9497	0.9123	27.4092	0.9419	29.3330	0.9294	log-Gaussian	0.6620	NA	0.1794	0.0550	0.0012	0.0568	-0.2374	-0.0052	-0.0112	0.2649	0.0744	0.0740	0.0819	0.0004	0.3643	0.1550	0.1981	0.0837	0.0733	0.0653	0.0073	0.2165	0.1151	0.0692	0.0911	0.0019	0.2076	0.1997	0.0859	0.0602	0.1301	0.0010	0.5097	0.1644	0.1944	0.0795	0.0766	0.0396	0.0118	0.0953	0.2157	0.8071	0.6559	0.4171	0.2051	0.6994	0.3517	0.8912	0.4681	0.3791
NA	K05813	0.0000	0.9298	0.2910	22.0576	20.8965	74.6228	21.5483	0.9474	22.0428	0.9070	17.7679	0.9176	Gaussian	0.7585	NA	-2.5400	0.7731	0.0012	-0.1577	-0.4793	-0.0865	0.9991	-2.2420	-2.9084	1.0440	1.1559	0.0327	0.0125	-2.6175	-2.4804	1.1768	1.0305	0.0270	0.0168	-3.0950	-1.5762	0.9724	1.2800	0.0016	0.2193	-2.4246	-3.0733	0.8539	1.8437	0.0049	0.0967	-1.8901	-3.1542	1.1119	1.0713	0.0903	0.0035	0.0953	0.6943	0.6698	0.5210	0.4602	0.1920	0.7067	0.4715	0.7633	0.5731	0.3202
NA	K05967	0.0000	0.8187	1.5082	17.4080	15.5252	52.6908	14.2290	0.7895	15.2921	0.8140	17.6809	0.8824	log-Gaussian	0.5191	NA	0.1658	0.0508	0.0012	0.0426	-0.0877	0.0014	-0.0196	0.2979	0.0036	0.0676	0.0749	0.0000	0.9616	0.2027	0.1375	0.0772	0.0676	0.0091	0.0430	0.2099	0.0893	0.0638	0.0840	0.0011	0.2886	0.1514	0.2337	0.0561	0.1211	0.0074	0.0547	0.2195	0.1163	0.0731	0.0705	0.0029	0.1002	0.0953	0.0353	0.9959	0.4351	0.5816	0.1620	0.7581	0.5283	0.7204	0.3073	0.5920
NA	K07473	0.0000	1.0000	89.8945	383.3194	373.5610	1047.4800	385.1940	1.0000	365.1540	1.0000	369.5050	1.0000	Gaussian	0.7561	NA	-25.2270	7.7028	0.0012	12.4225	25.5730	0.0670	1.0102	-17.2517	-35.0419	10.3747	11.4868	0.0976	0.0025	-12.0463	-35.3349	11.6740	10.2222	0.3031	0.0006	-32.6436	-12.4027	9.6690	12.7267	0.0008	0.3307	-23.5510	-33.2434	8.4678	18.2824	0.0058	0.0702	-12.0514	-37.2665	10.9791	10.5773	0.2734	0.0005	0.0953	0.7321	0.3982	0.8456	0.2937	0.1410	0.7771	0.5020	0.7426	0.7677	0.2275
Pyruvate	K14155	0.0000	1.0000	62.2066	177.4559	174.6795	290.3540	178.3600	1.0000	171.8095	1.0000	171.8740	1.0000	Gaussian	0.5836	NA	-9.4576	2.8744	0.0011	1.3248	-5.1135	-0.1308	1.8782	-12.8344	-5.3189	3.8782	4.2946	0.0011	0.2166	-12.2066	-7.3483	4.3726	3.8297	0.0056	0.0561	-8.2958	-11.4712	3.6212	4.7664	0.0227	0.0168	-9.2142	-10.5559	3.1563	6.8170	0.0038	0.1227	-13.1567	-6.0485	4.1379	3.9889	0.0016	0.1306	0.0953	0.3059	0.7660	0.4072	0.6324	0.4414	0.4153	0.4434	0.7927	0.2889	0.6270
NA	K19802	0.0000	0.7515	1.5922	13.9702	12.8162	35.3218	12.9433	0.7661	14.0819	0.7558	11.9373	0.7176	log-Gaussian	0.6519	NA	-0.1233	0.0377	0.0012	0.0084	0.0085	-0.0033	0.0149	-0.1349	-0.1090	0.0510	0.0565	0.0087	0.0546	-0.0997	-0.1413	0.0572	0.0501	0.0824	0.0051	-0.1675	-0.0463	0.0471	0.0620	0.0004	0.4558	-0.1202	-0.1369	0.0416	0.0898	0.0042	0.1287	-0.2038	-0.0484	0.0540	0.0521	0.0002	0.3532	0.0953	0.6399	0.7149	0.6938	0.4072	0.1038	0.8468	0.4454	0.7950	0.1778	0.8013
NA	K21576	0.0000	0.1316	1.0041	6.8332	6.3030	22.8655	6.6680	0.1637	6.2896	0.1279	5.6721	0.0706	Gaussian	0.7193	NA	-0.9199	0.2797	0.0011	-0.1717	-0.0263	0.0278	0.2366	-0.9223	-0.9160	0.3784	0.4190	0.0155	0.0297	-1.1736	-0.7254	0.4254	0.3725	0.0062	0.0526	-1.1708	-0.4850	0.3515	0.4626	0.0010	0.2955	-0.9864	-0.6063	0.3089	0.6670	0.0016	0.3642	-0.7175	-1.1051	0.4042	0.3894	0.0770	0.0049	0.0953	0.6698	0.6849	0.4072	0.6204	0.1512	0.7605	0.3947	0.8572	0.5467	0.3341
NA	K03711	0.0000	0.9708	2.7271	30.8006	27.2751	113.6430	28.1134	0.9825	26.3340	0.9651	24.0219	0.9529	log-Gaussian	0.6716	NA	-0.1306	0.0402	0.0013	0.0349	-0.0326	-0.0014	0.0433	-0.1230	-0.1402	0.0542	0.0600	0.0242	0.0203	-0.1642	-0.1048	0.0612	0.0536	0.0078	0.0518	-0.1433	-0.1084	0.0507	0.0668	0.0051	0.1056	-0.1243	-0.1601	0.0445	0.0960	0.0056	0.0966	-0.0972	-0.1612	0.0581	0.0560	0.0959	0.0043	0.0993	0.6849	0.6846	0.4214	0.6165	0.3266	0.6119	0.5001	0.7633	0.5836	0.3308
NA	K10670	0.0000	0.9152	3.7061	21.0458	19.5811	69.8346	20.7008	0.9298	18.8160	0.9186	16.7254	0.8824	log-Gaussian	0.7299	NA	-0.1147	0.0354	0.0013	0.0013	0.0016	0.0040	0.0283	-0.1317	-0.0937	0.0479	0.0530	0.0063	0.0780	-0.1634	-0.0773	0.0537	0.0470	0.0026	0.1014	-0.1280	-0.0919	0.0444	0.0584	0.0042	0.1168	-0.1203	-0.0881	0.0390	0.0843	0.0023	0.2972	-0.1310	-0.0995	0.0512	0.0493	0.0111	0.0446	0.0993	0.6063	0.7231	0.4004	0.7234	0.3171	0.6175	0.4325	0.8442	0.3779	0.4779

Pathway Results

Overview of the numbers of significant genes

Code

data.frame("what"  = c("No. of pathways that didn't have low abundance",
                       "No. of significant genes (fresh intervention)",
                       "No. of significant genes (past. intervention)"),
           "value" = c(nrow(dat_PWresults),
                       sum(dat_PWresults$freshInt_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_pvalue_correct < 0.1, na.rm = TRUE))) %>% 
  kable() %>% 
  kable_styling()

what	value
No. of pathways that didn't have low abundance	100
No. of significant genes (fresh intervention)	0
No. of significant genes (past. intervention)	0

… alternatively, looking at the raw p-values, not corrected for multiple testing

Code

data.frame("what"  = c("No. of pathways that didn't have low abundance",
                       "No. of significant genes (fresh intervention)",
                       "No. of significant genes (past. intervention)"),
           "value" = c(nrow(dat_PWresults),
                       sum(dat_PWresults$freshInt_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_pvalue < 0.05, na.rm = TRUE))) %>% 
  kable() %>% 
  kable_styling()

what	value
No. of pathways that didn't have low abundance	100
No. of significant genes (fresh intervention)	0
No. of significant genes (past. intervention)	41

Overview which specific genes are significant (with the multiple testing correction)

Code

dat_PWresults %>% 
  filter((freshInt_pvalue_correct < 0.1) |
           (pastInt_pvalue_correct < 0.1)) %>% 
  mutate(across(contains("pvalue_correct"), function(x) { ifelse(x < 0.1, x, "not significant") })) %>% 
  dplyr::select(pathway, contains("pvalue_correct")) %>% 
  arrange(freshInt_pvalue_correct, pastInt_pvalue_correct) %>% 
  kable() %>% 
  kable_styling()

pathway	freshInt_pvalue_correct	pastInt_pvalue_correct	freshInt_ageLow_pvalue_correct	pastInt_ageLow_pvalue_correct	freshInt_ageHigh_pvalue_correct	pastInt_ageHigh_pvalue_correct	freshInt_sexM_pvalue_correct	pastInt_sexM_pvalue_correct	freshInt_sexW_pvalue_correct	pastInt_sexW_pvalue_correct	freshInt_bmiLow_pvalue_correct	pastInt_bmiLow_pvalue_correct	freshInt_bmiHigh_pvalue_correct	pastInt_bmiHigh_pvalue_correct	freshInt_fibLow_pvalue_correct	pastInt_fibLow_pvalue_correct	freshInt_fibHigh_pvalue_correct	pastInt_fibHigh_pvalue_correct	freshInt_divLow_pvalue_correct	pastInt_divLow_pvalue_correct	freshInt_divHigh_pvalue_correct	pastInt_divHigh_pvalue_correct

Overview which specific genes are significant (without multiple testing correction)

Code

dat_PWresults %>% 
  filter((freshInt_pvalue < 0.05) |
           (pastInt_pvalue < 0.05)) %>% 
  dplyr::select(-contains("correct")) %>% 
  mutate(across(contains("pvalue"), function(x) { ifelse(x < 0.1, x, "not significant") })) %>% 
  dplyr::select(pathway, contains("pvalue")) %>% 
  arrange(freshInt_pvalue, pastInt_pvalue) %>% 
  kable() %>% 
  kable_styling()

pathway	freshInt_pvalue	pastInt_pvalue	freshInt_ageLow_pvalue	freshInt_ageHigh_pvalue	freshInt_sexM_pvalue	freshInt_sexW_pvalue	freshInt_bmiLow_pvalue	freshInt_bmiHigh_pvalue	freshInt_fibLow_pvalue	freshInt_fibHigh_pvalue	freshInt_divLow_pvalue	freshInt_divHigh_pvalue	pastInt_ageLow_pvalue	pastInt_ageHigh_pvalue	pastInt_sexM_pvalue	pastInt_sexW_pvalue	pastInt_bmiLow_pvalue	pastInt_bmiHigh_pvalue	pastInt_fibLow_pvalue	pastInt_fibHigh_pvalue	pastInt_divLow_pvalue	pastInt_divHigh_pvalue
D-Glutamine and D-glutamate metabolism	0.0637	0.0020	not significant	0.0336	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0086	0.0844	0.0047	0.0988	0.0045	not significant	0.009	0.0837	not significant	0.0044
Quorum sensing	0.0944	0.0417	not significant	0.0766	not significant	not significant	0.0125	not significant	not significant	not significant	not significant	not significant	not significant	0.0815	0.084	not significant	0.0243	not significant	0.0742	not significant	not significant	0.016
Streptomycin biosynthesis	0.0956	0.0488	not significant	not significant	not significant	0.0932	not significant	not significant	0.0334	not significant	not significant	not significant	0.0893	not significant	not significant	not significant	0.0287	not significant	not significant	0.0989	not significant	not significant
Arabinogalactan biosynthesis - Mycobacterium	not significant	0.0011	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0043	0.0856	0.0049	0.0597	0.0099	0.0446	0.0029	not significant	0.0403	0.0101
Glyoxylate and dicarboxylate metabolism	not significant	0.0031	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0126	0.0985	0.0616	0.0216	0.0019	not significant	0.0042	not significant	0.0361	0.0369
Histidine metabolism	not significant	0.0048	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0089	not significant	0.0196	0.0884	0.0059	not significant	0.0051	not significant	0.0243	0.0815
Alanine, aspartate and glutamate metabolism	not significant	0.0056	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0165	not significant	0.0289	0.0765	0.0055	not significant	0.0106	not significant	0.0736	0.0336
Sphingolipid metabolism	not significant	0.0058	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0009	not significant	not significant	0.0095	0.0073	not significant	0.0044	not significant	0.0129	not significant
Phenylpropanoid biosynthesis	not significant	0.0060	not significant	not significant	not significant	0.0978	not significant	not significant	not significant	not significant	not significant	not significant	0.0102	not significant	not significant	0.0222	0.014	not significant	0.006	not significant	0.0064	not significant
Starch and sucrose metabolism	not significant	0.0063	not significant	0.0773	not significant	not significant	0.0252	not significant	not significant	not significant	not significant	not significant	0.0194	not significant	0.0233	not significant	0.0103	not significant	0.0222	not significant	not significant	0.0221
Amino sugar and nucleotide sugar metabolism	not significant	0.0067	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0178	not significant	0.0589	0.051	0.0101	not significant	0.0135	not significant	0.0471	0.0639
Arginine and proline metabolism	not significant	0.0085	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0341	not significant	0.0801	0.0492	0.0044	not significant	0.0155	not significant	0.0721	0.0548
Glycerophospholipid metabolism	not significant	0.0099	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0134	0.0209	not significant	0.0122	not significant	0.0369	not significant	not significant	0.0163
Purine metabolism	not significant	0.0120	not significant	0.0883	not significant	not significant	0.0686	not significant	not significant	not significant	not significant	not significant	0.0724	0.0784	0.0208	not significant	0.0138	not significant	0.034	not significant	not significant	0.0143
Glycine, serine and threonine metabolism	not significant	0.0120	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0327	not significant	0.0267	not significant	0.0085	not significant	0.0216	not significant	not significant	0.0423
Arginine biosynthesis	not significant	0.0145	not significant	not significant	not significant	not significant	0.0878	not significant	not significant	not significant	not significant	not significant	0.0533	not significant	0.031	not significant	0.0082	not significant	0.0316	not significant	not significant	0.0111
Nicotinate and nicotinamide metabolism	not significant	0.0145	not significant	0.0712	not significant	not significant	0.0562	not significant	not significant	not significant	not significant	not significant	0.0747	0.0942	0.0603	not significant	0.0282	not significant	0.0364	not significant	not significant	0.0272
unnamed	not significant	0.0151	not significant	not significant	not significant	not significant	not significant	not significant	0.0843	not significant	not significant	not significant	0.0669	not significant	not significant	0.0644	0.008	not significant	0.0429	not significant	0.0296	not significant
Biotin metabolism	not significant	0.0186	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0453	not significant	0.0352	not significant	0.02	not significant	0.0463	not significant	0.0561	not significant
Lipoic acid metabolism	not significant	0.0224	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0259	not significant	not significant	0.0558	0.028	not significant	0.0274	not significant	0.0469	not significant
Porphyrin and chlorophyll metabolism	not significant	0.0253	not significant	not significant	not significant	not significant	0.0703	not significant	not significant	not significant	0.0741	not significant	0.086	not significant	0.044	not significant	0.0067	not significant	0.0517	not significant	not significant	0.0465
Nitrogen metabolism	not significant	0.0253	not significant	0.0752	not significant	not significant	0.0651	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0082	not significant	0.0583	not significant	not significant	0.035
Biofilm formation - Vibrio cholerae	not significant	0.0264	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0957	not significant	0.0296	0.0192	not significant	0.049	not significant	not significant	0.0397
Lysine biosynthesis	not significant	0.0287	not significant	0.0926	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.095	not significant	0.0243	not significant	0.0408	not significant	0.0661	not significant	not significant	0.051
RNA degradation	not significant	0.0293	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0674	not significant	0.0252	not significant	0.0421	not significant	0.0653	not significant	not significant	0.0356
Other glycan degradation	not significant	0.0301	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0097	not significant	not significant	0.0297	0.04	not significant	0.0254	not significant	0.0188	not significant
Glutathione metabolism	not significant	0.0308	not significant	0.042	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0972	not significant	not significant	not significant	0.0147	not significant	0.0682	not significant	not significant	not significant
DNA replication	not significant	0.0319	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0928	not significant	0.004	not significant	0.0507	not significant	0.0682	not significant	not significant	0.0105
Terpenoid backbone biosynthesis	not significant	0.0320	not significant	0.0815	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0677	not significant	0.0741	not significant	0.0361	not significant	0.0514	not significant	not significant	0.0501
Protein export	not significant	0.0352	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.083	not significant	0.0095	not significant	0.052	not significant	0.0621	not significant	not significant	0.0208
Selenocompound metabolism	not significant	0.0353	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0851	not significant	0.0665	not significant	0.0626	not significant	0.0703	not significant	not significant	0.0715
Peptidoglycan biosynthesis	not significant	0.0364	not significant	0.0829	not significant	not significant	0.0712	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.006	not significant	0.0633	not significant	0.0735	not significant	not significant	0.0072
Nucleotide excision repair	not significant	0.0384	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0957	not significant	0.0117	not significant	0.0741	not significant	0.0803	not significant	not significant	0.01
Thiamine metabolism	not significant	0.0391	not significant	0.0783	not significant	not significant	0.0521	not significant	not significant	not significant	not significant	not significant	0.0954	not significant	0.0595	not significant	0.0419	not significant	0.0691	not significant	not significant	0.0247
Galactose metabolism	not significant	0.0395	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0895	not significant	not significant	not significant	0.0916	not significant	0.0791	not significant	not significant	not significant
Phenylalanine metabolism	not significant	0.0401	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.079	0.0041	not significant	0.0604	not significant	not significant	0.0654
Aminoacyl-tRNA biosynthesis	not significant	0.0424	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0059	not significant	0.0689	not significant	0.0783	not significant	not significant	0.0126
Pantothenate and CoA biosynthesis	not significant	0.0426	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0289	not significant	0.0766	not significant	not significant	0.0576
Valine, leucine and isoleucine degradation	not significant	0.0447	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0693	not significant	not significant	not significant	not significant	0.0936	0.0481	not significant	0.0697	not significant	0.0568	not significant
Cell cycle - Caulobacter	not significant	0.0451	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0084	not significant	not significant	not significant	0.0956	not significant	not significant	0.0192
Glycerolipid metabolism	not significant	0.0484	not significant	not significant	not significant	not significant	0.0519	not significant	not significant	not significant	not significant	not significant	not significant	not significant	0.0245	not significant	0.0295	not significant	not significant	not significant	not significant	0.0081

Figure for publication

Code

# prepare the base dataset for the plot
dat_PWsignif <- dat_PWresults %>% 
  filter((freshInt_pvalue < 0.05) | (pastInt_pvalue < 0.05))
dat_plot <- data.frame(pathway      = rep(dat_PWsignif$pathway, times = 2),
                       baseline_medianTPMAbd = rep(dat_PWsignif$baseline_medianTPMAbd, times = 2),
                       model        = rep(dat_PWsignif$model, times = 2),
                       intervention = rep(c("Fresh intervention", "Pasteurized intervention"), each = nrow(dat_PWsignif)),
                       effect       = c(dat_PWsignif$freshInt_effect, dat_PWsignif$pastInt_effect),
                       pvalue       = c(dat_PWsignif$freshInt_pvalue, dat_PWsignif$pastInt_pvalue),
                       pastEffect_forSorting = rep(dat_PWsignif$pastInt_effect, times = 2)) %>% 
  dplyr::left_join(dat_PW %>% select(taxonomy_level3, taxonomy_level1) %>% distinct(taxonomy_level3, .keep_all = TRUE), by = c("pathway" = "taxonomy_level3")) %>% 
  arrange(desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((pvalue < 0.05) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (pvalue < 0.05) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (pvalue < 0.05) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (pvalue < 0.05) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model == "Gaussian") & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                   (model == "Gaussian") & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                   (model == "Gaussian") & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                   (model == "Gaussian") & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                   (model == "Gaussian") & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                   (model == "Gaussian") & (effect >= 0)        ~ "+ <0.0001 TPM",
                                   (model == "Gaussian") & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                   (model == "Gaussian") & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                   (model == "Gaussian") & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                   (model == "Gaussian") & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                   (model == "Gaussian") & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                   (model == "Gaussian") & (effect < 0)                   ~ "- >-0.0001 TPM",
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model == "log-Gaussian") & (effect >= 0)                                ~ "+ <0.01%",
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model == "log-Gaussian") & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model == "log-Gaussian") & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$pathway)), function(pw) {
  dat_pw <- dat_plot %>% filter(pathway == pw)
  dat_pw %>% 
    dplyr::bind_rows(data.frame(taxonomy_level1 = dat_pw$taxonomy_level1[1],
                                pathway       = pw,
                                intervention  = "Baseline median TPM abd.",
                                pvalue_signif = "",
                                effect_label  = case_when(dat_pw$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                          dat_pw$baseline_medianTPMAbd < .001  ~ paste0(round(dat_pw$baseline_medianTPMAbd, 4), " TPM"),
                                                          dat_pw$baseline_medianTPMAbd < .01   ~ paste0(round(dat_pw$baseline_medianTPMAbd, 3), " TPM"),
                                                          dat_pw$baseline_medianTPMAbd < .1    ~ paste0(round(dat_pw$baseline_medianTPMAbd, 2), " TPM"),
                                                          dat_pw$baseline_medianTPMAbd < 1     ~ paste0(round(dat_pw$baseline_medianTPMAbd, 1), " TPM"),
                                                          TRUE ~ paste0(round(dat_pw$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(pathway         = factor(pathway, levels = rev(unique(pathway))),
         intervention    = factor(intervention, levels = c("Baseline median TPM abd.", "Fresh intervention", "Pasteurized intervention")),
         pvalue_signif   = factor(pvalue_signif, levels = c("not significant",
                                                            "significant pos. effect (fresh intervention)",
                                                            "significant neg. effect (fresh intervention)",
                                                            "significant pos. effect (pasteurized intervention)",
                                                            "significant neg. effect (pasteurized intervention)",
                                                            "")),
         taxonomy_level1 = case_when(taxonomy_level1 == "Cellular Processes"             ~ "CP",
                                     taxonomy_level1 == "Genetic Information Processing" ~ "GIP",
                                     TRUE                                                ~ taxonomy_level1),
         taxonomy_level1 = factor(taxonomy_level1, levels = c("CP", "GIP", "Metabolism")))


# plot
dat_plot %>% 
  ggplot(aes(x = intervention, y = pathway)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("uncorrected p-value", values = c("not significant" = "gray95",
                                                      "significant pos. effect (fresh intervention)" = "#7cbde9",
                                                      "significant neg. effect (fresh intervention)" = "#1F78B4",
                                                      "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                                      "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(taxonomy_level1), scales = "free", space = "free") +
  ggtitle("Intervention effects on pathways") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"))

Code

# ggsave("FigureS17_pathwayScreening_pvalue.png", width = 10, height = 10)

Results for interaction effects

Overview of the numbers of significant genes

Code

data.frame("what"  = c("No. of pathways that didn't have low abundance",
                       "No. of significant pathways (fresh intervention - age low)",
                       "No. of significant pathways (fresh intervention - age high)",
                       "No. of significant pathways (past. intervention - age low)",
                       "No. of significant pathways (past. intervention - age high)",
                       "No. of significant pathways (fresh intervention - gender male)",
                       "No. of significant pathways (fresh intervention - gender female)",
                       "No. of significant pathways (past. intervention - gender male)",
                       "No. of significant pathways (past. intervention - gender female)",
                       "No. of significant pathways (fresh intervention - BMI low)",
                       "No. of significant pathways (fresh intervention - BMI high)",
                       "No. of significant pathways (past. intervention - BMI low)",
                       "No. of significant pathways (past. intervention - BMI high)",
                       "No. of significant pathways (fresh intervention - fiber low)",
                       "No. of significant pathways (fresh intervention - fiber high)",
                       "No. of significant pathways (past. intervention - fiber low)",
                       "No. of significant pathways (past. intervention - fiber high)",
                       "No. of significant pathways (fresh intervention - baseline div. low)",
                       "No. of significant pathways (fresh intervention - baseline div. high)",
                       "No. of significant pathways (past. intervention - baseline div. low)",
                       "No. of significant pathways (past. intervention - baseline div. high)"),
           "value" = c(nrow(dat_PWresults),
                       sum(dat_PWresults$freshInt_ageLow_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_ageHigh_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_ageLow_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_ageHigh_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_sexM_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_sexW_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_sexM_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_sexW_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_bmiLow_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_bmiHigh_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_bmiLow_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_bmiHigh_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_fibLow_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_fibHigh_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_fibLow_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_fibHigh_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_divLow_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_divHigh_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_divLow_pvalue_correct < 0.1, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_divHigh_pvalue_correct < 0.1, na.rm = TRUE))) %>% 
  kable() %>% 
  kable_styling()

what	value
No. of pathways that didn't have low abundance	100
No. of significant pathways (fresh intervention - age low)	0
No. of significant pathways (fresh intervention - age high)	0
No. of significant pathways (past. intervention - age low)	0
No. of significant pathways (past. intervention - age high)	0
No. of significant pathways (fresh intervention - gender male)	0
No. of significant pathways (fresh intervention - gender female)	0
No. of significant pathways (past. intervention - gender male)	0
No. of significant pathways (past. intervention - gender female)	0
No. of significant pathways (fresh intervention - BMI low)	0
No. of significant pathways (fresh intervention - BMI high)	0
No. of significant pathways (past. intervention - BMI low)	0
No. of significant pathways (past. intervention - BMI high)	0
No. of significant pathways (fresh intervention - fiber low)	0
No. of significant pathways (fresh intervention - fiber high)	0
No. of significant pathways (past. intervention - fiber low)	0
No. of significant pathways (past. intervention - fiber high)	0
No. of significant pathways (fresh intervention - baseline div. low)	0
No. of significant pathways (fresh intervention - baseline div. high)	0
No. of significant pathways (past. intervention - baseline div. low)	0
No. of significant pathways (past. intervention - baseline div. high)	0

… alternatively looking at the uncorrected p-values

Code

data.frame("what"  = c("No. of pathways that didn't have low abundance",
                       "No. of significant pathways (fresh intervention - age low)",
                       "No. of significant pathways (fresh intervention - age high)",
                       "No. of significant pathways (past. intervention - age low)",
                       "No. of significant pathways (past. intervention - age high)",
                       "No. of significant pathways (fresh intervention - gender male)",
                       "No. of significant pathways (fresh intervention - gender female)",
                       "No. of significant pathways (past. intervention - gender male)",
                       "No. of significant pathways (past. intervention - gender female)",
                       "No. of significant pathways (fresh intervention - BMI low)",
                       "No. of significant pathways (fresh intervention - BMI high)",
                       "No. of significant pathways (past. intervention - BMI low)",
                       "No. of significant pathways (past. intervention - BMI high)",
                       "No. of significant pathways (fresh intervention - fiber low)",
                       "No. of significant pathways (fresh intervention - fiber high)",
                       "No. of significant pathways (past. intervention - fiber low)",
                       "No. of significant pathways (past. intervention - fiber high)",
                       "No. of significant pathways (fresh intervention - baseline div. low)",
                       "No. of significant pathways (fresh intervention - baseline div. high)",
                       "No. of significant pathways (past. intervention - baseline div. low)",
                       "No. of significant pathways (past. intervention - baseline div. high)"),
           "value" = c(nrow(dat_PWresults),
                       sum(dat_PWresults$freshInt_ageLow_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_ageHigh_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_ageLow_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_ageHigh_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_sexM_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_sexW_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_sexM_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_sexW_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_bmiLow_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_bmiHigh_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_bmiLow_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_bmiHigh_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_fibLow_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_fibHigh_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_fibLow_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_fibHigh_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_divLow_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$freshInt_divHigh_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_divLow_pvalue < 0.05, na.rm = TRUE),
                       sum(dat_PWresults$pastInt_divHigh_pvalue < 0.05, na.rm = TRUE))) %>% 
  kable() %>% 
  kable_styling()

what	value
No. of pathways that didn't have low abundance	100
No. of significant pathways (fresh intervention - age low)	0
No. of significant pathways (fresh intervention - age high)	3
No. of significant pathways (past. intervention - age low)	17
No. of significant pathways (past. intervention - age high)	2
No. of significant pathways (fresh intervention - gender male)	0
No. of significant pathways (fresh intervention - gender female)	0
No. of significant pathways (past. intervention - gender male)	28
No. of significant pathways (past. intervention - gender female)	7
No. of significant pathways (fresh intervention - BMI low)	6
No. of significant pathways (fresh intervention - BMI high)	1
No. of significant pathways (past. intervention - BMI low)	36
No. of significant pathways (past. intervention - BMI high)	1
No. of significant pathways (fresh intervention - fiber low)	1
No. of significant pathways (fresh intervention - fiber high)	1
No. of significant pathways (past. intervention - fiber low)	20
No. of significant pathways (past. intervention - fiber high)	1
No. of significant pathways (fresh intervention - baseline div. low)	2
No. of significant pathways (fresh intervention - baseline div. high)	0
No. of significant pathways (past. intervention - baseline div. low)	10
No. of significant pathways (past. intervention - baseline div. high)	30

baseline diversity

Code

# prepare the base dataset for the plot
dat_signif <- dat_PWresults %>% 
  filter((freshInt_divLow_pvalue < 0.05) | (freshInt_divHigh_pvalue < 0.05) |
           (pastInt_divLow_pvalue < 0.05) | (pastInt_divHigh_pvalue < 0.05))
dat_plot <- data.frame(pathway       = rep(dat_signif$pathway,  times = 4),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(baseline div. low)", "Fresh intervention\n(baseline div. high)",
                                             "Pasteurized intervention\n(baseline div. low)", "Pasteurized intervention\n(baseline div. high)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_divLow_effect, dat_signif$freshInt_divHigh_effect,
                                         dat_signif$pastInt_divLow_effect,  dat_signif$pastInt_divHigh_effect),
                       pvalue        = c(dat_signif$freshInt_divLow_pvalue, dat_signif$freshInt_divHigh_pvalue,
                                         dat_signif$pastInt_divLow_pvalue,  dat_signif$pastInt_divHigh_pvalue),
                       pastEffect_forSorting = rep(dat_signif$pastInt_divLow_effect, times = 4)) %>% 
  dplyr::left_join(dat_PW %>% select(taxonomy_level3, taxonomy_level1) %>% distinct(taxonomy_level3, .keep_all = TRUE), by = c("pathway" = "taxonomy_level3")) %>% 
  arrange(desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((pvalue < 0.05) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (pvalue < 0.05) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (pvalue < 0.05) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (pvalue < 0.05) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 TPM",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 TPM",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$pathway)), function(pw) {
  dat_pw <- dat_plot %>% filter(pathway == pw)
  dat_pw %>% 
    dplyr::bind_rows(data.frame(taxonomy_level1 = dat_pw$taxonomy_level1[1],
                                pathway         = pw,
                                intervention    = "Baseline median TPM abd.",
                                pvalue_signif   = "",
                                effect_label    = case_when(dat_pw$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                            dat_pw$baseline_medianTPMAbd < .001  ~ paste0(round(dat_pw$baseline_medianTPMAbd, 4), " TPM"),
                                                            dat_pw$baseline_medianTPMAbd < .01   ~ paste0(round(dat_pw$baseline_medianTPMAbd, 3), " TPM"),
                                                            dat_pw$baseline_medianTPMAbd < .1    ~ paste0(round(dat_pw$baseline_medianTPMAbd, 2), " TPM"),
                                                            dat_pw$baseline_medianTPMAbd < 1     ~ paste0(round(dat_pw$baseline_medianTPMAbd, 1), " TPM"),
                                                            TRUE ~ paste0(round(dat_pw$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(pathway         = factor(pathway, levels = rev(unique(pathway))),
         intervention    = gsub(intervention, pattern = "low",  replacement = "[2.8, 3.85)"),
         intervention    = gsub(intervention, pattern = "high", replacement = "[3.85, 4.4)"),
         intervention    = factor(intervention, levels = c("Baseline median TPM abd.",
                                                           "Fresh intervention\n(baseline div. [2.8, 3.85))",
                                                           "Fresh intervention\n(baseline div. [3.85, 4.4))",
                                                           "Pasteurized intervention\n(baseline div. [2.8, 3.85))",
                                                           "Pasteurized intervention\n(baseline div. [3.85, 4.4))")),
         pvalue_signif   = factor(pvalue_signif, levels = c("not significant",
                                                            "significant pos. effect (fresh intervention)",
                                                            "significant neg. effect (fresh intervention)",
                                                            "significant pos. effect (pasteurized intervention)",
                                                            "significant neg. effect (pasteurized intervention)",
                                                            "")),
         taxonomy_level1 = case_when(taxonomy_level1 == "Cellular Processes"             ~ "CP",
                                     taxonomy_level1 == "Genetic Information Processing" ~ "GIP",
                                     TRUE                                                ~ taxonomy_level1),
         taxonomy_level1 = factor(taxonomy_level1, levels = c("CP", "GIP", "Metabolism")))

# plot
dat_plot %>% 
  ggplot(aes(x = intervention, y = pathway)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("uncorrected p-value", values = c("not significant" = "gray95",
                                                      "significant pos. effect (fresh intervention)" = "#7cbde9",
                                                      "significant neg. effect (fresh intervention)" = "#1F78B4",
                                                      "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                                      "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(taxonomy_level1), scales = "free", space = "free") +
  ggtitle("Intervention effects on pathways") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"))

age

Code

# prepare the base dataset for the plot
dat_signif <- dat_PWresults %>% 
  filter((freshInt_ageLow_pvalue < 0.05) | (freshInt_ageHigh_pvalue < 0.05) |
           (pastInt_ageLow_pvalue < 0.05) | (pastInt_ageHigh_pvalue < 0.05))
dat_plot <- data.frame(pathway       = rep(dat_signif$pathway,  times = 4),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(age low)", "Fresh intervention\n(age high)",
                                             "Pasteurized intervention\n(age low)", "Pasteurized intervention\n(age high)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_ageLow_effect, dat_signif$freshInt_ageHigh_effect,
                                         dat_signif$pastInt_ageLow_effect,  dat_signif$pastInt_ageHigh_effect),
                       pvalue        = c(dat_signif$freshInt_ageLow_pvalue, dat_signif$freshInt_ageHigh_pvalue,
                                         dat_signif$pastInt_ageLow_pvalue,  dat_signif$pastInt_ageHigh_pvalue),
                       pastEffect_forSorting = rep(dat_signif$pastInt_ageLow_effect, times = 4)) %>% 
  dplyr::left_join(dat_PW %>% select(taxonomy_level3, taxonomy_level1) %>% distinct(taxonomy_level3, .keep_all = TRUE), by = c("pathway" = "taxonomy_level3")) %>% 
  arrange(desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((pvalue < 0.05) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (pvalue < 0.05) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (pvalue < 0.05) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (pvalue < 0.05) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 TPM",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 TPM",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$pathway)), function(pw) {
  dat_pw <- dat_plot %>% filter(pathway == pw)
  dat_pw %>% 
    dplyr::bind_rows(data.frame(taxonomy_level1 = dat_pw$taxonomy_level1[1],
                                pathway         = pw,
                                intervention    = "Baseline median TPM abd.",
                                pvalue_signif   = "",
                                effect_label    = case_when(dat_pw$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                            dat_pw$baseline_medianTPMAbd < .001  ~ paste0(round(dat_pw$baseline_medianTPMAbd, 4), " TPM"),
                                                            dat_pw$baseline_medianTPMAbd < .01   ~ paste0(round(dat_pw$baseline_medianTPMAbd, 3), " TPM"),
                                                            dat_pw$baseline_medianTPMAbd < .1    ~ paste0(round(dat_pw$baseline_medianTPMAbd, 2), " TPM"),
                                                            dat_pw$baseline_medianTPMAbd < 1     ~ paste0(round(dat_pw$baseline_medianTPMAbd, 1), " TPM"),
                                                            TRUE ~ paste0(round(dat_pw$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(pathway         = factor(pathway, levels = rev(unique(pathway))),
         intervention    = gsub(intervention, pattern = "age low",  replacement = "age 21-49"),
         intervention    = gsub(intervention, pattern = "age high", replacement = "age 50-69"),
         intervention    = factor(intervention, levels = c("Baseline median TPM abd.",
                                                           "Fresh intervention\n(age 21-49)",
                                                           "Fresh intervention\n(age 50-69)",
                                                           "Pasteurized intervention\n(age 21-49)",
                                                           "Pasteurized intervention\n(age 50-69)")),
         pvalue_signif   = factor(pvalue_signif, levels = c("not significant",
                                                            "significant pos. effect (fresh intervention)",
                                                            "significant neg. effect (fresh intervention)",
                                                            "significant pos. effect (pasteurized intervention)",
                                                            "significant neg. effect (pasteurized intervention)",
                                                            "")),
         taxonomy_level1 = case_when(taxonomy_level1 == "Cellular Processes"             ~ "CP",
                                     taxonomy_level1 == "Genetic Information Processing" ~ "GIP",
                                     TRUE                                                ~ taxonomy_level1),
         taxonomy_level1 = factor(taxonomy_level1, levels = c("CP", "GIP", "Metabolism")))

# plot
dat_plot %>% 
  ggplot(aes(x = intervention, y = pathway)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("uncorrected p-value", values = c("not significant" = "gray95",
                                                      "significant pos. effect (fresh intervention)" = "#7cbde9",
                                                      "significant neg. effect (fresh intervention)" = "#1F78B4",
                                                      "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                                      "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(taxonomy_level1), scales = "free", space = "free") +
  ggtitle("Intervention effects on pathways") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"))

sex

Code

# prepare the base dataset for the plot
dat_signif <- dat_PWresults %>% 
  filter((freshInt_sexM_pvalue < 0.05) | (freshInt_sexW_pvalue < 0.05) |
           (pastInt_sexM_pvalue < 0.05) | (pastInt_sexW_pvalue < 0.05))
dat_plot <- data.frame(pathway       = rep(dat_signif$pathway,  times = 4),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(gender male)", "Fresh intervention\n(gender female)",
                                             "Pasteurized intervention\n(gender male)", "Pasteurized intervention\n(gender female)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_sexM_effect, dat_signif$freshInt_sexW_effect,
                                         dat_signif$pastInt_sexM_effect,  dat_signif$pastInt_sexW_effect),
                       pvalue        = c(dat_signif$freshInt_sexM_pvalue, dat_signif$freshInt_sexW_pvalue,
                                         dat_signif$pastInt_sexM_pvalue,  dat_signif$pastInt_sexW_pvalue),
                       pastEffect_forSorting = rep(dat_signif$pastInt_sexM_effect, times = 4)) %>% 
  dplyr::left_join(dat_PW %>% select(taxonomy_level3, taxonomy_level1) %>% distinct(taxonomy_level3, .keep_all = TRUE), by = c("pathway" = "taxonomy_level3")) %>% 
  arrange(desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((pvalue < 0.05) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (pvalue < 0.05) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (pvalue < 0.05) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (pvalue < 0.05) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 TPM",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 TPM",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$pathway)), function(pw) {
  dat_pw <- dat_plot %>% filter(pathway == pw)
  dat_pw %>% 
    dplyr::bind_rows(data.frame(taxonomy_level1 = dat_pw$taxonomy_level1[1],
                                pathway         = pw,
                                intervention    = "Baseline median TPM abd.",
                                pvalue_signif   = "",
                                effect_label    = case_when(dat_pw$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                            dat_pw$baseline_medianTPMAbd < .001  ~ paste0(round(dat_pw$baseline_medianTPMAbd, 4), " TPM"),
                                                            dat_pw$baseline_medianTPMAbd < .01   ~ paste0(round(dat_pw$baseline_medianTPMAbd, 3), " TPM"),
                                                            dat_pw$baseline_medianTPMAbd < .1    ~ paste0(round(dat_pw$baseline_medianTPMAbd, 2), " TPM"),
                                                            dat_pw$baseline_medianTPMAbd < 1     ~ paste0(round(dat_pw$baseline_medianTPMAbd, 1), " TPM"),
                                                            TRUE ~ paste0(round(dat_pw$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(pathway         = factor(pathway, levels = rev(unique(pathway))),
         intervention    = gsub(intervention, pattern = "gender ", replacement = ""),
         intervention    = factor(intervention, levels = c("Baseline median TPM abd.",
                                                           "Fresh intervention\n(male)",
                                                           "Fresh intervention\n(female)",
                                                           "Pasteurized intervention\n(male)",
                                                           "Pasteurized intervention\n(female)")),
         pvalue_signif   = factor(pvalue_signif, levels = c("not significant",
                                                            "significant pos. effect (fresh intervention)",
                                                            "significant neg. effect (fresh intervention)",
                                                            "significant pos. effect (pasteurized intervention)",
                                                            "significant neg. effect (pasteurized intervention)",
                                                            "")),
         taxonomy_level1 = case_when(taxonomy_level1 == "Cellular Processes" ~ "CP",
                                     TRUE                                    ~ taxonomy_level1),
         taxonomy_level1 = factor(taxonomy_level1, levels = c("CP", "Genetic Information Processing", "Metabolism")))

# plot
dat_plot %>% 
  ggplot(aes(x = intervention, y = pathway)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("uncorrected p-value", values = c("not significant" = "gray95",
                                                      "significant pos. effect (fresh intervention)" = "#7cbde9",
                                                      "significant neg. effect (fresh intervention)" = "#1F78B4",
                                                      "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                                      "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(taxonomy_level1), scales = "free", space = "free") +
  ggtitle("Intervention effects on pathways") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"))

BMI

Code

# prepare the base dataset for the plot
dat_signif <- dat_PWresults %>% 
  filter((freshInt_bmiLow_pvalue < 0.05) | (freshInt_bmiHigh_pvalue < 0.05) |
           (pastInt_bmiLow_pvalue < 0.05) | (pastInt_bmiHigh_pvalue < 0.05))
dat_plot <- data.frame(pathway       = rep(dat_signif$pathway,  times = 4),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(BMI low)", "Fresh intervention\n(BMI high)",
                                             "Pasteurized intervention\n(BMI low)", "Pasteurized intervention\n(BMI high)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_bmiLow_effect, dat_signif$freshInt_bmiHigh_effect,
                                         dat_signif$pastInt_bmiLow_effect,  dat_signif$pastInt_bmiHigh_effect),
                       pvalue        = c(dat_signif$freshInt_bmiLow_pvalue, dat_signif$freshInt_bmiHigh_pvalue,
                                         dat_signif$pastInt_bmiLow_pvalue,  dat_signif$pastInt_bmiHigh_pvalue),
                       pastEffect_forSorting = rep(dat_signif$pastInt_bmiLow_effect, times = 4)) %>% 
  dplyr::left_join(dat_PW %>% select(taxonomy_level3, taxonomy_level1) %>% distinct(taxonomy_level3, .keep_all = TRUE), by = c("pathway" = "taxonomy_level3")) %>% 
  arrange(desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((pvalue < 0.05) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (pvalue < 0.05) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (pvalue < 0.05) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (pvalue < 0.05) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 TPM",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 TPM",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$pathway)), function(pw) {
  dat_pw <- dat_plot %>% filter(pathway == pw)
  dat_pw %>% 
    dplyr::bind_rows(data.frame(taxonomy_level1 = dat_pw$taxonomy_level1[1],
                                pathway         = pw,
                                intervention    = "Baseline median TPM abd.",
                                pvalue_signif   = "",
                                effect_label    = case_when(dat_pw$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                            dat_pw$baseline_medianTPMAbd < .001  ~ paste0(round(dat_pw$baseline_medianTPMAbd, 4), " TPM"),
                                                            dat_pw$baseline_medianTPMAbd < .01   ~ paste0(round(dat_pw$baseline_medianTPMAbd, 3), " TPM"),
                                                            dat_pw$baseline_medianTPMAbd < .1    ~ paste0(round(dat_pw$baseline_medianTPMAbd, 2), " TPM"),
                                                            dat_pw$baseline_medianTPMAbd < 1     ~ paste0(round(dat_pw$baseline_medianTPMAbd, 1), " TPM"),
                                                            TRUE ~ paste0(round(dat_pw$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(pathway         = factor(pathway, levels = rev(unique(pathway))),
         intervention    = gsub(intervention, pattern = "low",  replacement = "[18, 25)"),
         intervention    = gsub(intervention, pattern = "high", replacement = "[25, 31]"),
         intervention    = factor(intervention, levels = c("Baseline median TPM abd.",
                                                           "Fresh intervention\n(BMI [18, 25))",
                                                           "Fresh intervention\n(BMI [25, 31])",
                                                           "Pasteurized intervention\n(BMI [18, 25))",
                                                           "Pasteurized intervention\n(BMI [25, 31])")),
         pvalue_signif   = factor(pvalue_signif, levels = c("not significant",
                                                            "significant pos. effect (fresh intervention)",
                                                            "significant neg. effect (fresh intervention)",
                                                            "significant pos. effect (pasteurized intervention)",
                                                            "significant neg. effect (pasteurized intervention)",
                                                            "")),
         taxonomy_level1 = case_when(taxonomy_level1 == "Cellular Processes"             ~ "CP",
                                     taxonomy_level1 == "Genetic Information Processing" ~ "GIP",
                                     TRUE                                                ~ taxonomy_level1),
         taxonomy_level1 = factor(taxonomy_level1, levels = c("CP", "GIP", "Metabolism")))

# plot
dat_plot %>% 
  ggplot(aes(x = intervention, y = pathway)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("uncorrected p-value", values = c("not significant" = "gray95",
                                                      "significant pos. effect (fresh intervention)" = "#7cbde9",
                                                      "significant neg. effect (fresh intervention)" = "#1F78B4",
                                                      "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                                      "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(taxonomy_level1), scales = "free", space = "free") +
  ggtitle("Intervention effects on pathways") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"))

fiber

Code

# prepare the base dataset for the plot
dat_signif <- dat_PWresults %>% 
  filter((freshInt_fibLow_pvalue < 0.05) | (freshInt_fibHigh_pvalue < 0.05) |
           (pastInt_fibLow_pvalue < 0.05) | (pastInt_fibHigh_pvalue < 0.05))
dat_plot <- data.frame(pathway       = rep(dat_signif$pathway,  times = 4),
                       baseline_medianTPMAbd = rep(dat_signif$baseline_medianTPMAbd, times = 4),
                       model         = rep(dat_signif$model, times = 4),
                       intervention  = rep(c("Fresh intervention\n(fiber low)", "Fresh intervention\n(fiber high)",
                                             "Pasteurized intervention\n(fiber low)", "Pasteurized intervention\n(fiber high)"), each = nrow(dat_signif)),
                       effect        = c(dat_signif$freshInt_fibLow_effect, dat_signif$freshInt_fibHigh_effect,
                                         dat_signif$pastInt_fibLow_effect,  dat_signif$pastInt_fibHigh_effect),
                       pvalue        = c(dat_signif$freshInt_fibLow_pvalue, dat_signif$freshInt_fibHigh_pvalue,
                                         dat_signif$pastInt_fibLow_pvalue,  dat_signif$pastInt_fibHigh_pvalue),
                       pastEffect_forSorting = rep(dat_signif$pastInt_fibLow_effect, times = 4)) %>% 
  dplyr::left_join(dat_PW %>% select(taxonomy_level3, taxonomy_level1) %>% distinct(taxonomy_level3, .keep_all = TRUE), by = c("pathway" = "taxonomy_level3")) %>% 
  arrange(desc(pastEffect_forSorting)) %>% 
  mutate(pvalue_signif = case_when((pvalue < 0.05) & grepl("Fresh", intervention) & (effect > 0) ~ "significant pos. effect (fresh intervention)",
                                   (pvalue < 0.05) & grepl("Fresh", intervention) & (effect < 0) ~ "significant neg. effect (fresh intervention)",
                                   (pvalue < 0.05) & (effect > 0) ~ "significant pos. effect (pasteurized intervention)",
                                   (pvalue < 0.05) & (effect < 0) ~ "significant neg. effect (pasteurized intervention)",
                                   TRUE          ~ "not significant"),
         effect_label  = case_when((model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 1)        ~ paste0("+", round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.1)      ~ paste0("+", round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.01)     ~ paste0("+", round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.001)    ~ paste0("+", round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0.0001)   ~ paste0("+", round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect >= 0)        ~ "+ <0.0001 TPM",
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -1)       ~ paste0(round(effect, 0), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.1)     ~ paste0(round(effect, 1), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.01)    ~ paste0(round(effect, 2), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.001)   ~ paste0(round(effect, 3), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect <= -0.0001)  ~ paste0(round(effect, 4), " TPM"),
                                   (model %in% c("Gaussian", "ZI Gaussian")) & (effect < 0)                   ~ "- >-0.0001 TPM",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 1)             ~ paste0("+", round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.1)           ~ paste0("+", round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) >= 0.01)          ~ paste0("+", round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect >= 0)                                ~ "+ <0.01%",
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -1)            ~ paste0(round((100 * (exp(effect) - 1)), 0), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.1)          ~ paste0(round((100 * (exp(effect) - 1)), 1), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & ((100 * (exp(effect) - 1)) <= -0.01)         ~ paste0(round((100 * (exp(effect) - 1)), 2), "%"),
                                   (model %in% c("log-Gaussian", "ZI log-Gaussian")) & (effect < 0)                                 ~ "- >-0.01%"))

# artificially add the baseline rel. abd. as an 'intervention' and 'effect_label' to be able to nicely plot it
dat_plot_list <- lapply(as.character(unique(dat_plot$pathway)), function(pw) {
  dat_pw <- dat_plot %>% filter(pathway == pw)
  dat_pw %>% 
    dplyr::bind_rows(data.frame(taxonomy_level1 = dat_pw$taxonomy_level1[1],
                                pathway         = pw,
                                intervention    = "Baseline median TPM abd.",
                                pvalue_signif   = "",
                                effect_label    = case_when(dat_pw$baseline_medianTPMAbd < .0001 ~ "<0.0001 TPM",
                                                            dat_pw$baseline_medianTPMAbd < .001  ~ paste0(round(dat_pw$baseline_medianTPMAbd, 4), " TPM"),
                                                            dat_pw$baseline_medianTPMAbd < .01   ~ paste0(round(dat_pw$baseline_medianTPMAbd, 3), " TPM"),
                                                            dat_pw$baseline_medianTPMAbd < .1    ~ paste0(round(dat_pw$baseline_medianTPMAbd, 2), " TPM"),
                                                            dat_pw$baseline_medianTPMAbd < 1     ~ paste0(round(dat_pw$baseline_medianTPMAbd, 1), " TPM"),
                                                            TRUE ~ paste0(round(dat_pw$baseline_medianTPMAbd, 0), " TPM"))))
})
dat_plot <- dat_plot_list %>% dplyr::bind_rows() %>% 
  mutate(pathway         = factor(pathway, levels = rev(unique(pathway))),
         # intervention    = gsub(intervention, pattern = "low",  replacement = "[18, 25)"),
         # intervention    = gsub(intervention, pattern = "high", replacement = "[25, 31]"),
         intervention    = factor(intervention, levels = c("Baseline median TPM abd.",
                                                           "Fresh intervention\n(fiber low)",
                                                           "Fresh intervention\n(fiber high)",
                                                           "Pasteurized intervention\n(fiber low)",
                                                           "Pasteurized intervention\n(fiber high)")),
         pvalue_signif   = factor(pvalue_signif, levels = c("not significant",
                                                            "significant pos. effect (fresh intervention)",
                                                            "significant neg. effect (fresh intervention)",
                                                            "significant pos. effect (pasteurized intervention)",
                                                            "significant neg. effect (pasteurized intervention)",
                                                            "")),
         taxonomy_level1 = case_when(taxonomy_level1 == "Cellular Processes"             ~ "CP",
                                     taxonomy_level1 == "Genetic Information Processing" ~ "GIP",
                                     TRUE                                                ~ taxonomy_level1),
         taxonomy_level1 = factor(taxonomy_level1, levels = c("CP", "GIP", "Metabolism")))

# plot
dat_plot %>% 
  ggplot(aes(x = intervention, y = pathway)) +
  geom_tile(aes(fill = pvalue_signif)) +
  geom_text(aes(label = effect_label)) +
  scale_fill_manual("uncorrected p-value", values = c("not significant" = "gray95",
                                                      "significant pos. effect (fresh intervention)" = "#7cbde9",
                                                      "significant neg. effect (fresh intervention)" = "#1F78B4",
                                                      "significant pos. effect (pasteurized intervention)" = "#ffb366",
                                                      "significant neg. effect (pasteurized intervention)" = "#FF7F00"),
                    na.value = "white") +
  facet_grid(rows = vars(taxonomy_level1), scales = "free", space = "free") +
  ggtitle("Intervention effects on pathways") +
  theme(axis.title = element_blank(),
        panel.grid = element_blank(),
        strip.background = element_rect(fill = "gray97", color = "gray97"))

Joint plot of condensed information

Focus on the four biggest phylum groups: Firmicutes, Bacteroidetes, Actinobacteria and Proteobacteria

Code

# data preparation
plot_dat_wide <- dat_PWresults %>% 
  dplyr::left_join(dat_PW %>% select(taxonomy_level1, taxonomy_level3) %>% distinct(taxonomy_level3, .keep_all = T),
                   by = c("pathway" = "taxonomy_level3")) %>% 
  group_by(taxonomy_level1) %>% 
  dplyr::summarize(n_pathways                   = n(),
                   n_freshSignifEffects_overall = sum(freshInt_pvalue         < 0.05),
                   n_freshSignifEffects_divLow  = sum(freshInt_divLow_pvalue  < 0.05),
                   n_freshSignifEffects_divHigh = sum(freshInt_divHigh_pvalue < 0.05),
                   n_freshSignifEffects_ageLow  = sum(freshInt_ageLow_pvalue  < 0.05),
                   n_freshSignifEffects_ageHigh = sum(freshInt_ageHigh_pvalue < 0.05),
                   n_freshSignifEffects_sexM    = sum(freshInt_sexM_pvalue    < 0.05),
                   n_freshSignifEffects_sexW    = sum(freshInt_sexW_pvalue    < 0.05),
                   n_freshSignifEffects_bmiLow  = sum(freshInt_bmiLow_pvalue  < 0.05),
                   n_freshSignifEffects_bmiHigh = sum(freshInt_bmiHigh_pvalue < 0.05),
                   n_freshSignifEffects_fibLow  = sum(freshInt_fibLow_pvalue  < 0.05),
                   n_freshSignifEffects_fibHigh = sum(freshInt_fibHigh_pvalue < 0.05),
                   n_pastSignifEffects_overall  = sum(pastInt_pvalue          < 0.05),
                   n_pastSignifEffects_divLow   = sum(pastInt_divLow_pvalue   < 0.05),
                   n_pastSignifEffects_divHigh  = sum(pastInt_divHigh_pvalue  < 0.05),
                   n_pastSignifEffects_ageLow   = sum(pastInt_ageLow_pvalue   < 0.05),
                   n_pastSignifEffects_ageHigh  = sum(pastInt_ageHigh_pvalue  < 0.05),
                   n_pastSignifEffects_sexM     = sum(pastInt_sexM_pvalue     < 0.05),
                   n_pastSignifEffects_sexW     = sum(pastInt_sexW_pvalue     < 0.05),
                   n_pastSignifEffects_bmiLow   = sum(pastInt_bmiLow_pvalue   < 0.05),
                   n_pastSignifEffects_bmiHigh  = sum(pastInt_bmiHigh_pvalue  < 0.05),
                   n_pastSignifEffects_fibLow   = sum(pastInt_fibLow_pvalue   < 0.05),
                   n_pastSignifEffects_fibHigh  = sum(pastInt_fibHigh_pvalue  < 0.05))


# reshape dataset to long format
plot_dat_long <- data.frame(taxonomy_level1      = rep(plot_dat_wide$taxonomy_level1, times = sum(grepl("freshSignifEffects", colnames(plot_dat_wide)))),
                            n_pathways           = rep(plot_dat_wide$n_pathways, times = sum(grepl("freshSignifEffects", colnames(plot_dat_wide)))),
                            effect_group         = rep(c("overall", "divInt", "divInt", "ageInt", "ageInt", "sexInt", "sexInt", "bmiInt", "bmiInt", "fibInt", "fibInt"),
                                                       each = nrow(plot_dat_wide)),
                            effect_type          = rep(c("overall", "divLow", "divHigh", "ageLow", "ageHigh", "sexM", "sexW", "bmiLow", "bmiHigh", "fibLow", "fibHigh"),
                                                       each = nrow(plot_dat_wide)),
                            n_freshSignifEffects = c(plot_dat_wide$n_freshSignifEffects_overall,
                                                     plot_dat_wide$n_freshSignifEffects_divLow,
                                                     plot_dat_wide$n_freshSignifEffects_divHigh,
                                                     plot_dat_wide$n_freshSignifEffects_ageLow,
                                                     plot_dat_wide$n_freshSignifEffects_ageHigh,
                                                     plot_dat_wide$n_freshSignifEffects_sexM,
                                                     plot_dat_wide$n_freshSignifEffects_sexW,
                                                     plot_dat_wide$n_freshSignifEffects_bmiLow,
                                                     plot_dat_wide$n_freshSignifEffects_bmiHigh,
                                                     plot_dat_wide$n_freshSignifEffects_fibLow,
                                                     plot_dat_wide$n_freshSignifEffects_fibHigh),
                            n_pastSignifEffects  = c(plot_dat_wide$n_pastSignifEffects_overall,
                                                     plot_dat_wide$n_pastSignifEffects_divLow,
                                                     plot_dat_wide$n_pastSignifEffects_divHigh,
                                                     plot_dat_wide$n_pastSignifEffects_ageLow,
                                                     plot_dat_wide$n_pastSignifEffects_ageHigh,
                                                     plot_dat_wide$n_pastSignifEffects_sexM,
                                                     plot_dat_wide$n_pastSignifEffects_sexW,
                                                     plot_dat_wide$n_pastSignifEffects_bmiLow,
                                                     plot_dat_wide$n_pastSignifEffects_bmiHigh,
                                                     plot_dat_wide$n_pastSignifEffects_fibLow,
                                                     plot_dat_wide$n_pastSignifEffects_fibHigh)) %>% 
  mutate(taxLevel1_label = paste0(taxonomy_level1, "\n", n_pathways, " pathways"),
         effect_group    = factor(effect_group, levels = c("overall", "divInt", "ageInt", "sexInt", "bmiInt", "fibInt")),
         effect_type     = factor(effect_type, levels = rev(c("overall", "divLow", "divHigh", "ageLow", "ageHigh", "sexM", "sexW", "bmiLow", "bmiHigh", "fibLow", "fibHigh"))))

# make the dataset even longer
plot_dat_longer <- data.frame(taxonomy_level1 = rep(plot_dat_long$taxonomy_level1, times = 2),
                              n_pathways      = rep(plot_dat_long$n_pathways,      times = 2),
                              taxLevel1_label = rep(plot_dat_long$taxLevel1_label, times = 2),
                              effect_group    = rep(plot_dat_long$effect_group,    times = 2),
                              effect_type     = rep(plot_dat_long$effect_type,     times = 2),
                              intervention    = rep(c("Fresh", "Past."), each = nrow(plot_dat_long)),
                              parameter       = "n_signifEffects",
                              value           = c(plot_dat_long$n_freshSignifEffects,
                                                  plot_dat_long$n_pastSignifEffects)) %>% 
  group_by(taxonomy_level1, parameter, effect_group, intervention) %>% 
  mutate(alpha_value = case_when(effect_group == "overall"  ~ 0.8,
                                 value == min(value)        ~ 0,
                                 TRUE                       ~ 1)) %>% 
  ungroup() %>% 
  mutate(fill_tiles = case_when(effect_type == "overall" ~ "no",
                                TRUE ~ taxonomy_level1),
         alpha_tiles = case_when(effect_type %in% c("divLow", "ageLow", "sexM", "bmiLow", "fibLow") ~ 0.05,
                                 TRUE ~ 0.15))
plot_dat_longer <- plot_dat_longer %>%
  mutate(taxLevel1_label = factor(taxLevel1_label, levels = plot_dat_longer %>% arrange(desc(n_pathways)) %>% 
                                    pull(taxLevel1_label) %>% unique()),
         effect_type     = case_when(effect_type == "divLow"  ~ "div. [2.8, 3.85)",
                                     effect_type == "divHigh" ~ "div. [3.85, 4.4)",
                                     effect_type == "ageLow"  ~ "age 21-49",
                                     effect_type == "ageHigh" ~ "age 50-69",
                                     effect_type == "sexM"    ~ "male",
                                     effect_type == "sexW"    ~ "female",
                                     effect_type == "bmiLow"  ~ "BMI [18, 25)",
                                     effect_type == "bmiHigh" ~ "BMI [25, 31]",
                                     effect_type == "fibLow"  ~ "fiber (8, 30)",
                                     effect_type == "fibHigh" ~ "fiber [30, 58)",
                                     TRUE                     ~ effect_type),
         effect_type     = factor(effect_type, levels = c("div. [3.85, 4.4)", "div. [2.8, 3.85)",
                                                          "BMI [25, 31]", "BMI [18, 25)",
                                                          "female", "male",
                                                          "age 50-69", "age 21-49",
                                                          "fiber [30, 58)", "fiber (8, 30)", "overall")))


# create a color vector for color coding the interaction terms
col_vector <- RColorBrewer::brewer.pal(3, name = "Dark2")

# plot
ggplot(plot_dat_longer, aes(x = intervention, y = effect_type)) +
  geom_tile(aes(fill = fill_tiles, alpha = alpha_tiles)) +
  geom_text(aes(label = value, alpha = alpha_value)) +
  facet_grid(effect_group ~ taxLevel1_label, scales = "free") +
  scale_fill_manual(values = c("no"                             = "white",
                               "Metabolism"                     = col_vector[1],
                               "Genetic Information Processing" = col_vector[2],
                               "Cellular Processes"             = col_vector[3])) +
  theme(axis.title      = element_blank(),
        panel.grid      = element_blank(),
        legend.position = "none",
        strip.background.x = element_rect(fill = "gray97", color = "gray97"),
        strip.text.y    = element_blank())